JP2000157297A - Method for detecting factor of kawasaki disease - Google Patents
Method for detecting factor of kawasaki diseaseInfo
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、川崎病の発症に関
与する川崎病因子の検出法に関する。The present invention relates to a method for detecting a Kawasaki disease factor involved in the development of Kawasaki disease.
【０００２】[0002]
【従来技術と発明が解決しようとする課題】川崎病は、
１９６３年川崎冨作により初めて報告された乳幼児期に
認められる病気であり、近年では、人種毎の発症頻度が
異なるものの世界各地で発症例が報告されている。川崎
病は男児に好発し、その患者年齢としては４才以下が80
〜85％を占めているが、年長者にも発症が認められてい
る。また、冠動脈障害や冠動脈瘤による突然死をきたす
こともある。死亡頻度は、0.3〜0.5％であるが、乳幼児
における心臓疾患の主要原因でもある。また、後遺症と
して冠状動脈瘤及び狭窄を来すことがあり、心筋梗塞の
原因となる。発症初期におけるガンマグロブリン療法等
により冠動脈瘤形成頻度の減少が認められるものの、ガ
ンマグロブリン療法の無効例も多く存在している。川崎
病はその病態が、ブドウ球菌由来の細菌外毒素（TSST−
1：スーパー抗原の一種）による毒素性ショック症候群
（Toxic shock Syndrom: TSS）等に見られる全身性の症
状に類似しており、リンパ球、マクロファージ系の細胞
活性化を初めとした免疫学的変化顕著である（臨床免
疫,26(7): 738-744, 1994）。更には、患者の急性期末
梢血リンパ球においてＶβ２及びＶβ８陽性Ｔ細胞が選
択的に増加しているとの報告もあり（J., Kotozin, B.
L., Jujo, K., Melish, M.E., Glode, M.P., Kohsaka,
T. and Leung, D.Y.M. (1992) Selective expansion of
T cells expressing T-cell receptor variable region
s V beta 2 and V beta8 in Kawasaki disease, Proc N
atl Acad Sci USA 89, 4066; Abe J., Kotozin,B.L., M
eissner, C., Melish, M.E., Masato, T., Fuluton,
D., Romagne, F.,Malissen, B. and Leung, D.Y.M. (19
93) Characterization of T cell repertoire changes
in acute Kawasaki disease, J Exp Med 177,791; Curt
is, N., Zheng, R., Lamb, J.R. and Levin, M.(1995)
Evidence for superantigen mediated process in Kawa
saki disease, Arch Dis Child 72, 308; Donald et. a
l.,1995, J. Immunol. (1995), 155: 5018-5021)、その
発症にはスーパー抗原(SA)の関与が示唆されていた。2. Description of the Related Art Kawasaki disease
This disease was first reported by Tosaku Kawasaki in 1963, and was observed in infancy. In recent years, cases of occurrence have been reported in various parts of the world, although the frequency of occurrence varies depending on the race. Kawasaki disease is common among boys, and the age of the patient is 80 years old or younger.
It accounts for ~ 85%, but has also been found in older people. In addition, sudden death due to coronary artery disorder or coronary aneurysm may occur. Death frequency is 0.3-0.5%, but is also the leading cause of heart disease in infants. Further, as a sequela, coronary aneurysm and stenosis may occur, which causes myocardial infarction. Although the frequency of coronary aneurysm formation is reduced by gamma globulin therapy and the like in the early stage of onset, there are many cases where gamma globulin therapy is ineffective. Kawasaki disease is characterized by a bacterial exotoxin (TSST-
1: a kind of superantigen), similar to systemic symptoms such as Toxic shock Syndrom (TSS), and immunological changes including lymphocyte and macrophage cell activation It is remarkable (clinical immunity, 26 (7): 738-744, 1994). Furthermore, it has been reported that Vβ2 and Vβ8 positive T cells are selectively increased in acute peripheral blood lymphocytes of patients (J., Kotozin, B. et al.
L., Jujo, K., Melish, ME, Glode, MP, Kohsaka,
T. and Leung, DYM (1992) Selective expansion of
T cells expressing T-cell receptor variable region
s V beta 2 and V beta 8 in Kawasaki disease, Proc N
atl Acad Sci USA 89, 4066; Abe J., Kotozin, BL, M
eissner, C., Melish, ME, Masato, T., Fuluton,
D., Romagne, F., Malissen, B. and Leung, DYM (19
93) Characterization of T cell repertoire changes
in acute Kawasaki disease, J Exp Med 177,791; Curt
is, N., Zheng, R., Lamb, JR and Levin, M. (1995)
Evidence for superantigen mediated process in Kawa
saki disease, Arch Dis Child 72, 308; Donald et.a
l., 1995, J. Immunol. (1995), 155: 5018-5021), suggesting the involvement of superantigen (SA) in its onset.
【０００３】また、ガンマグロブリン療法が川崎病に奏
効するということも、川崎病の発症にスーパー抗原が関
与することを示唆している。即ち、ガンマグロブリン製
剤中にブドウ球菌由来スーパー抗原に対する抗体が存在
し、この抗SA抗体がSAによるＴ細胞活性化特異的に抑制
するとの報告（Takei, S., Arora, Y.K. and Walker,M.
(1993) Intravenous immunoglobulin contains specif
ic antibodies inhibitory to activation of T cell b
y staphylococcal toxin superantigens, J.Clin Inves
t, 91, 602-607)がある。しかしながら、その病因は未
だ解明されていない状態である。[0003] The fact that gamma globulin therapy responds to Kawasaki disease also suggests that superantigens are involved in the development of Kawasaki disease. That is, there is a report that an antibody against a staphylococcal superantigen is present in a gamma globulin preparation, and that this anti-SA antibody specifically suppresses T cell activation by SA (Takei, S., Arora, YK and Walker, M., et al.
(1993) Intravenous immunoglobulin contains specif
ic antibodies inhibitory to activation of T cell b
y staphylococcal toxin superantigens, J. Clin Inves
t, 91, 602-607). However, its etiology has not yet been elucidated.
【０００４】従来、本症の診断は、1) ５日以上続く発
熱；2) 四肢末端の変化；硬性浮腫、紅斑等；3) 不定形
発疹；4) 両眼球結膜の充血；5) ***、口腔所見；口腔
の紅潮、イチゴ舌、口腔咽頭粘膜のびまん性発赤；6)
急性期における非化膿性頚部リンパ節腫眼からなる臨床
症状の内、５つ以上の症状を伴うことを基準として行わ
れている。しかし、必ずしもこれら主要症状が早期に揃
って出現するとは限らず、特に年長発症の場合には、重
症例が多い上、主要症状の出現に時間がかかることか
ら、早期に的確な診断を行うための基準の確立が強く求
められている。Conventionally, this disease has been diagnosed by: 1) fever lasting 5 days or more; 2) changes in extremities; hard edema, erythema, etc .; 3) irregular rash; 4) conjunctival congestion in both eyes; Oral findings; flushing of the oral cavity, strawberry tongue, diffuse redness of the oropharyngeal mucosa; 6)
Non-suppurative cervical lymphadenopathy in the acute phase is performed on the basis of clinical symptoms involving five or more symptoms. However, these major symptoms do not always appear at an early stage, and especially in the case of older onset, an accurate diagnosis is made early because many severe cases and the appearance of major symptoms take time. There is a strong demand for establishing standards for this.
【０００５】[0005]
【課題を解決するための手段】本発明者らは、川崎病の
的確な早期診断を確立するために、鋭意研究を重ねた結
果、川崎病の発症期の患者に、川崎病に特異的な因子の
存在を示唆する、科学的に検出可能な徴候が現れている
ことを見出し、本発明を完成するに至った。即ち、本発
明は、Ｖβ６.５陽性Ｔ細胞数の増加、又はＶβ６.５及
びＶβ２.１陽性Ｔ細胞数の増加を指標とする、川崎病
因子の検出法を提供するものである。「川崎病因子」と
は、川崎病の臨床症状を顕在化させる因子を意味し、何
らかの原因で川崎病に罹患した患者の血中に存在する病
因物質を含む。そのような物質として、例えば、細菌性
スーパー抗原の可能性がある。スーパー抗原の代表例と
して、Ａ群連鎖球菌由来の外毒素（Streptococcal pyog
enic exptoxin C(SPEC))を挙げることができる。Means for Solving the Problems The present inventors have conducted intensive studies in order to establish an accurate early diagnosis of Kawasaki disease. The inventors have found that scientifically detectable signs suggesting the presence of the factor have appeared, and have completed the present invention. That is, the present invention provides a method for detecting Kawasaki disease factor using an increase in the number of Vβ6.5-positive T cells or an increase in the number of Vβ6.5 and Vβ2.1-positive T cells as an index. “Kawasaki disease factor” refers to a factor that manifests the clinical symptoms of Kawasaki disease, and includes an etiological substance present in the blood of a patient suffering from Kawasaki disease for some reason. Such a substance may be, for example, a bacterial superantigen. As a representative example of the superantigen, an exotoxin derived from group A streptococcus (Streptococcal pyog
enic exptoxin C (SPEC)).
【０００６】本発明の川崎病因子の検出法を実施するに
は、川崎病を疑われる患者から末梢血を得、ＴＣＲＶ領
域の遺伝子群を増幅し、ＴＣＲＶサブファミリーの発現
頻度を検出し、Ｖβ６.５陽性Ｔ細胞、又はＶβ６.５陽
性Ｔ細胞及びＶβ２.１陽性Ｔ細胞の増加を評価する。
Ｔ細胞のＶβレパートリーは、当該技術分野で既知の任
意の方法で増幅し、発現を調べることができる。例え
ば、患者から末梢血を得、トータルＲＮＡを抽出し、既
知のアダプター付加ＰＣＲ（アダプターライゲーション
ＰＣＲ＝ＡＬ−ＰＣＲ）でＴＣＲＶ領域の遺伝子群を増
幅し、リバース・ドット・ブロット法（Reverse dot bl
otting）により、ＴＣＲＶサブファミリーに特異的なオ
リゴプローブを用いて、それらサブファミリーの発現状
態を調べ、ＴＣＲＶβ６.５、又はＴＣＲＶβ６.５及び
ＴＣＲＶβ２.１サブファミリーの発現頻度の有意な増
大（絶対的及び/又は相対的）を比較する。リバース・
ドット・ブロット法に用いるオリゴヌクレオチドプロー
ブは、ＴＣＲＶβサブファミリーに特異的であり、少な
くとも、ＴＣＲＶβ２.１及びＴＣＲＶβ６.５に対応す
るプローブを含むことを条件として任意である。本発明
の実施に用いることができるオリゴヌクレオチドプロー
ブの一例として、表７〜１３に記載のオリゴヌクレオチ
ドプローブが挙げられる。これらは配列番号１０及び１
１に記載の塩基配列で示されるオリゴヌクレオチドプロ
ーブ（表１０のVB02-1及び表１１のVB06-4、それぞれＴ
ＣＲＶβ２.１及びＴＣＲＶβ６.５に対応する）と、配
列番号１２〜９６に記載のオリゴヌクレオチドを含む。
これらオリゴヌクレオチドプローブを用いる場合、最終
的に得られたサブファミリーの発現頻度のデータから、
配列番号１０及び１１に記載のオリゴヌクレオチドプロ
ーブに対応するＴＣＲＶβ６.５、又はＴＣＲＶβ６.５
及びＴＣＲＶβ２.１サブファミリーの発現頻度が他の
サブファミリーの発現頻度よりも有意に高いことを指標
として、川崎病因子の存在を決定すればよい。To carry out the method for detecting a Kawasaki disease factor of the present invention, peripheral blood is obtained from a patient suspected of having Kawasaki disease, the genes of the TCRV region are amplified, the expression frequency of the TCRV subfamily is detected, and Vβ6 Evaluate the increase of .5 positive T cells or Vβ6.5 positive T cells and Vβ2.1 positive T cells.
The V cell repertoire of T cells can be expanded and examined for expression by any method known in the art. For example, peripheral blood is obtained from a patient, total RNA is extracted, a group of genes in the TCRV region is amplified by a known adapter-added PCR (adapter ligation PCR = AL-PCR), and reverse dot blotting (Reverse dot bl
otting), using oligo probes specific for the TCRV subfamily to examine the expression status of those subfamilies and to significantly increase the frequency of expression of the TCRVβ6.5 or TCRVβ6.5 and TCRVβ2.1 subfamilies (absolute And / or relative). reverse·
The oligonucleotide probe used for the dot blot method is specific to the TCRVβ subfamily, and is optional provided that it contains at least probes corresponding to TCRVβ2.1 and TCRVβ6.5. Examples of the oligonucleotide probes that can be used in the practice of the present invention include the oligonucleotide probes described in Tables 7 to 13. These are SEQ ID NOs: 10 and 1.
The oligonucleotide probe represented by the nucleotide sequence shown in Table 1 (VB02-1 in Table 10 and VB06-4 in Table 11;
CRVβ2.1 and TCRVβ6.5) and the oligonucleotides set forth in SEQ ID NOs: 12-96.
When using these oligonucleotide probes, from the data on the expression frequency of the finally obtained subfamily,
TCRVβ6.5 or TCRVβ6.5 corresponding to the oligonucleotide probes of SEQ ID NOS: 10 and 11.
The presence of the Kawasaki disease factor may be determined by using the expression frequency of the TCRVβ2.1 subfamily and the expression frequency of the other subfamilies significantly higher as an index.
【０００７】別法として、Ｖβ２.１及びＶβ６.５に特
異的なプライマーで増幅し、その後、増幅量を定量して
もよい。定量法としては、電気泳動に供し、ハウスキー
ピング遺伝子をインナーコントロールとして定量するこ
とができる。川崎病因子の存在は、ＴＣＲＶβ鎖におけ
る、Ｖβ２.１及びＶβ６.５を検出し、Ｖβ２.１とＶ
β６.５陽性Ｔ細胞が、川崎病発症期において発現量の
増加が健常人群コントロールの平均値に比べ有意に高い
場合を陽性、低い場合を陰性とする。その他のファクタ
ー、例えば、Ｔ細胞の増加がポリクローナルであるこ
と、各種炎症性サイトカインの増加、従来の症状の発現
も診断に際して補助的に用いることができる。[0007] Alternatively, amplification may be carried out with primers specific for Vβ2.1 and Vβ6.5, and then the amount of amplification may be quantified. As a quantification method, housekeeping genes can be quantified as an inner control by subjecting to electrophoresis. The presence of Kawasaki disease factor detected Vβ2.1 and Vβ6.5 in the TCRVβ chain,
ββ-positive T cells are positive if the increase in the expression level during the Kawasaki disease onset phase is significantly higher than the average value of the control group of healthy individuals, and negative if the increase is low. Other factors such as polyclonal increase in T cells, increase in various inflammatory cytokines, and expression of conventional symptoms can also be used as an aid in diagnosis.
【０００８】実施例１ 表１に示す川崎病患者２２名の治療前（発症期)、治療
後（７９日以上経過後）及び発熱を伴う川崎病以外の疾
患群の子供１４名、及び健常群の大人１０名について末
梢血中のＴＣＲＶレパートリーを表２に示すオリゴヌク
レオチドプライマーを用いるアダプター付加ＰＣＲ法で
増幅し、表７〜１３に記載のオリゴヌクレオチドプロー
ブを用いるリバース・ドット・ブロット法でＴＣＲＶサ
ブファミリーの発現状態を調べ、ＴＣＲＶβ６.５及び
ＴＣＲＶβ２.１サブファミリーの発現頻度を解析し
た。 Ｉ． ＴＣＲＶ領域の増幅表１ 川崎病因子の検出対象 Example 1 Twenty-two Kawasaki disease patients shown in Table 1 before treatment (onset), after treatment (after 79 days or more), and 14 children of a disease group other than Kawasaki disease accompanied by fever, and a healthy group The TCRV repertoire in the peripheral blood of 10 adults was amplified by an adapter-added PCR method using the oligonucleotide primers shown in Table 2, and was subjected to reverse dot blotting using The expression status of the families was examined, and the expression frequencies of the TCRVβ6.5 and TCRVβ2.1 subfamilies were analyzed. I. Amplification of TCRV region Table 1 Targets for detection of Kawasaki disease factor
【表１】 [Table 1]
【０００９】表２ アダプター付加ＰＣＲ法及びＴＣＲ
αβcＤＮＡの増幅に用いるオリゴヌクレオチドプライ
マー Table 2 PCR method with adapter and TCR
Oligonucleotide primers used for amplification of αβ cDNA
【表２】 [Table 2]
【００１０】表２のＴＣＲαβｃＤＮＡのＰＣＲ増幅用
オリゴヌクレオチドプライマーの内、ＣＡ１（配列番号
４）、ＣＡ２（配列番号５）及びＣＡ３（配列番号６）
は２種類のＣα領域の共通部分に、ＣＢ１（配列番号
７）、ＣＢ２（配列番号８）及びＣＢ３（配列番号９）
は２種類のＣβ領域の共通配列上に、それぞれ対応する
よう設定されている。Among the oligonucleotide primers for PCR amplification of TCRαβ cDNA shown in Table 2, CA1 (SEQ ID NO: 4), CA2 (SEQ ID NO: 5) and CA3 (SEQ ID NO: 6)
Indicates that CB1 (SEQ ID NO: 7), CB2 (SEQ ID NO: 8) and CB3 (SEQ ID NO: 9)
Are set on the common sequence of the two types of Cβ regions so as to correspond to each other.
【００１１】（１）ｃＤＮＡの合成 表２に記載のＰＣＲプライマーＰ１０ＥＡ（配列番号
１）、Ｐ２０ＥＡ（配列番号２）及びＢＳＬ−１８（配
列番号３）を用いた。対象から末梢血５−１０mlを採取
し、Ficoll-Hypaque (ファルマシア社)を用いてＰＢＭ
Ｃと血清に分けた。Trizol-Reagents(BRL社)を使用し、
業者の指示に従って、ＰＢＭＣよりトータルＲＮＡを抽
出した。抽出したＲＮＡから、Adaptor−ligation ＰＣ
Ｒ法（Tsuruta, Y., Iwagami,S., Furue, S., Teraoka,
H., Yoshida, T., Sakata, T. and Suzuki, R. (1993)
Detection of human T cell receptor cDNAs (α、
β、γ、and δ）by ligation of a universal adaptor
to variable region J Immunol Methods 161, 7)によ
り、ＴＣＲ Ｖ領域を含む遺伝子群を増幅した。まず、
抽出されたＲＮＡから、ＢＳＬ−１８プライマー（表２
及び配列番号３）を用いて、１stストランドＤＮＡを合
成し、次いで、２ndストランドＤＮＡを合成し、フェノ
ール抽出し、次いでポリエチレングリコール（PEG)を用
いたＤＮＡ沈殿法で余分なＢＳＬ−１８を除去し、精製
した。(1) Synthesis of cDNA PCR primers P10EA (SEQ ID NO: 1), P20EA (SEQ ID NO: 2) and BSL-18 (SEQ ID NO: 3) described in Table 2 were used. 5-10 ml of peripheral blood is collected from the subject and subjected to PBM using Ficoll-Hypaque (Pharmacia).
C and serum. Using Trizol-Reagents (BRL),
According to the manufacturer's instructions, total RNA was extracted from PBMC. Adapter-ligation PC from extracted RNA
R method (Tsuruta, Y., Iwagami, S., Furue, S., Teraoka,
H., Yoshida, T., Sakata, T. and Suzuki, R. (1993)
Detection of human T cell receptor cDNAs (α,
β, γ, and δ) by ligation of a universal adaptor
The gene group containing the TCR V region was amplified by to variable region J Immunol Methods 161, 7). First,
From the extracted RNA, a BSL-18 primer (Table 2)
And the second strand DNA was synthesized by using SEQ ID NO: 3), then the second strand DNA was synthesized, extracted with phenol, and the extra BSL-18 was removed by a DNA precipitation method using polyethylene glycol (PEG). And purified.
【００１２】（２）アダプターの付加 １）Ｐ２０ＥＡ／Ｐ１０ＥＡアダプターの調製 RNaseを含有しない滅菌水(和光純薬）に溶かした各１０
０pmol／μｌのＰ２０ＥＡとＰ１０ＥＡ（配列番号２及
び１）を、１.５ｍｌスクリューキャップ付き滅菌チュ
ーブ内で１：１の割合で混合し、蓋をして７０℃で10分
間加熱し、室温まで徐冷した。得られたＰ２０ＥＡ／Ｐ
１０ＥＡ溶液を冷房室に移し10℃以下になった時点で−
70℃にて凍結させ、使用するまで−20℃で保存した。(2) Addition of Adapter 1) Preparation of P20EA / P10EA Adapter Each of 10 adapters dissolved in sterile water (Wako Pure Chemical Industries, Ltd.) containing no RNase.
0 pmol / μl of P20EA and P10EA (SEQ ID NOs: 2 and 1) were mixed at a ratio of 1: 1 in a sterile tube with a 1.5 ml screw cap, heated with a lid at 70 ° C. for 10 minutes, and gradually cooled to room temperature. Cooled down. P20EA / P obtained
Transfer the 10EA solution to the cooling room, and when the temperature drops below 10 ° C-
It was frozen at 70 ° C and stored at -20 ° C until use.
【００１３】２）２本鎖ｃＤＮＡに対するアダプターの
付加 氷上で、２本鎖ｃＤＮＡ溶液（12.5μｌ）、x5 T4 DNA
Ligase Buffer (BRL;250 mM Tris-HCl (pH 7.6), 50 mM
MgCl2, 5 mM ATP, 5 mM DTT,25%(W/V) PEG8000)（5.0
μl = x1 buffer)、50 pmol/μl P20EA/P10EA アダプ
ター(5.0μl,10μＭ）、Ｔ4 DNA Ligase (BRL) （2.5μ
l)を混合し、合計２５.０μlの反応液をピペッティング
及びタッピングで良く攪拌し、4℃でFlash遠心した後、
16℃で20時間靜置した。反応終了後、4℃でFlash遠心し
た。次いで、アダプターを付加した２本鎖ｃＤＮＡをフ
ェノール抽出で精製し、ＰＥＧを用いるＤＮＡ沈殿によ
り余分なＰ２０ＥＡ／Ｐ１０ＥＡアダプターを除去し
た。2) Addition of adapter to double-stranded cDNA On ice, double-stranded cDNA solution (12.5 μl), x5 T4 DNA
Ligase Buffer (BRL; 250 mM Tris-HCl (pH 7.6), 50 mM
MgCl 2 , 5 mM ATP, 5 mM DTT, 25% (W / V) PEG8000) (5.0
μl = x1 buffer), 50 pmol / μl P20EA / P10EA adapter (5.0 μl, 10 μM), T4 DNA Ligase (BRL) (2.5 μl
l) was mixed, and a total of 25.0 μl of the reaction solution was thoroughly stirred by pipetting and tapping, followed by Flash centrifugation at 4 ° C.
The mixture was allowed to stand at 16 ° C for 20 hours. After completion of the reaction, the mixture was subjected to Flash centrifugation at 4 ° C. Next, the double-stranded cDNA to which the adapter was added was purified by phenol extraction, and the extra P20EA / P10EA adapter was removed by DNA precipitation using PEG.
【００１４】３）ＴＣＲ遺伝子Ｃ領域側に付いたアダプ
ターの切断 上記２）で得た34.0μlのアダプター付加２本鎖ｃＤＮ
Ａに、氷上で、0.1% Triton-X(5.0μｌ, 0.01%, TAKAR
A）,0.1% BSA (5.0μl, 0.01%, TAKARA), x10 buffer H
(5.0 μｌ, x1 buffer, TAKARA)を混合し、最後に Not
I (約５０Ｕ／μｌ, TAKARA）を加えて、合計50.0μｌ
の混合液をタッピングにより良く混合し、4℃でFlash遠
心を行った後、37℃で2時間靜置した。反応終了後、直
ちに氷上に移した。次いで、NotI処理した2本鎖ｃＤＮ
Ａをフェノール抽出して精製し、ＰＥＧを用いるＤＮＡ
沈殿法でNotI処理により生じたＰ２０ＥＡ／Ｐ１０ＥＡ
アダプターを除去した。得られたＤＮＡを２０μｌの滅
菌水に懸濁し、‐20℃で保存した。3) Cleavage of the adapter attached to the C region of the TCR gene 34.0 μl of the adapter-added double-stranded cDN obtained in 2) above
A. On ice, 0.1% Triton-X (5.0 μl, 0.01%, TAKAR
A), 0.1% BSA (5.0μl, 0.01%, TAKARA), x10 buffer H
(5.0 μl, x1 buffer, TAKARA).
I (about 50 U / μl, TAKARA) and add 50.0 μl
Was mixed well by tapping, and the mixture was subjected to Flash centrifugation at 4 ° C, and then allowed to stand at 37 ° C for 2 hours. Immediately after the reaction was completed, the mixture was transferred to ice. Then, NotI-treated double-stranded cDN
A is purified by phenol extraction and DNA using PEG
P20EA / P10EA generated by NotI treatment by precipitation method
Adapter was removed. The obtained DNA was suspended in 20 μl of sterile water and stored at −20 ° C.
【００１５】（３）ＰＣＲ ＰＣＲは、以下の表に示すプライマーセットを用いて行
った。表３ ＰＣＲ用プライマーセット(3) PCR PCR was carried out using the primer sets shown in the following table. Table 3 Primer set for PCR
【表３】 [Table 3]
【００１６】１）第１回（１st）ＰＣＲ 上記（２）で最終的に２０μｌに調製したｃＤＮＡを0.
5ml滅菌チューブに１μlずつ分注し、氷上で保管した。
x10 ＰＣＲ用バッファー(Promega)、25mM MgCl2(Promeg
a)、P20EA/CA1及びP20EA/CB1プライマーセット（50pmol
/μl）、25mM dNTP mix(g)を氷上で解凍し、ボルテック
スで攪拌した後、Flash遠心により溶液を回収した。上
記の試薬を用い、α、β各500μl（10回分）の１stＰＣ
Ｒ溶液を氷上で以下の組成に従い調製した。1) First (1st) PCR The cDNA finally prepared in 20 μl in the above (2) was used in 0.1 μl.
1 μl was dispensed into a 5 ml sterile tube and stored on ice.
x10 PCR buffer (Promega), 25mM MgCl 2 (Promeg
a), P20EA / CA1 and P20EA / CB1 primer set (50 pmol
/ μl), 25 mM dNTP mix (g) was thawed on ice, vortexed, and the solution was recovered by Flash centrifugation. Using the above reagents, 500 µl each of α and β (10 times) of 1stPC
An R solution was prepared on ice according to the following composition.
【００１７】表４ α、β鎖用１stＰＣＲ溶液 Table 4 1st PCR solution for α and β chains
【表４】 混合物を良く攪拌し、Flash遠心したのち、α、β各４
９０μｌ(10回分）の1stＰＣＲ溶液を、先にｃＤＮＡを
分注しておいたチューブに４９μlずつ分注した。残っ
たＰＣＲ溶液は、ネガティブコントロールに用いた。
α、β各10本ずつのチューブにミネラルオイル（SIGM
A）約５０μlを分注し、合計１５０μlの混合液を用い
て以下の手順でＰＣＲ増幅を行った（変成94℃30秒を１
サイクル、変成94℃１分、アニール55℃、1.5分、伸長7
2℃、2分を２０サイクル、Last extension72℃、5分以
下で4℃で保存）。Flash遠心後、-70℃に靜置し、氷結
させた。室温に戻し、ミネラルオイルのみが解凍した時
点で、マイクロピペットによりこれを除去した。[Table 4] Stir the mixture well and centrifuge in Flash, then 4 each for α and β
90 μl (10 times) of the 1st PCR solution was dispensed in 49 μl portions into the tube into which cDNA was previously dispensed. The remaining PCR solution was used for a negative control.
Mineral oil (SIGM) in 10 tubes each for α and β
A) Approximately 50 μl was dispensed, and PCR amplification was performed using a total of 150 μl of the mixed solution according to the following procedure (denaturation was performed at 94 ° C. for 30 seconds for 1 second).
Cycle, denaturation 94 ° C for 1 minute, annealing 55 ° C, 1.5 minutes, extension 7
20 cycles of 2 minutes at 2 ° C, Last extension 72 ° C, stored at 4 ° C for 5 minutes or less). After flash centrifugation, the mixture was allowed to stand at -70 ° C and frozen. After returning to room temperature and only the mineral oil was thawed, it was removed with a micropipette.
【００１８】２）第２回(２nd）ＰＣＲ 上記１）で増幅したＰＣＲ産物を、0.5ml滅菌チューブ
に５μlずつ、ネガティブコントロールとして増幅した
産物は２μｌを分注し、氷上で保管した。x10 ＰＣＲ用
バッファー(Promega)、25mM MgCl2(Promega)、P20EA/CA
2及びP20EA/CB2プライマーセット（5pmol/μl）、25mM
dNTP mix(TOYOBO)を氷上で解凍し、ボルテックスで良く
攪拌した後、Flash遠心により溶液を回収した。上記の
試薬を用い、α、β各500μl（10回分）の２ndＰＣＲ溶
液を氷上で以下の組成に従い調製した。2) 2nd (2nd) PCR 5 μl of the PCR product amplified in the above 1) was dispensed into a 0.5 ml sterilized tube, and 2 μl of the amplified product was stored as a negative control and stored on ice. x10 PCR buffer (Promega), 25mM MgCl 2 (Promega), P20EA / CA
2 and P20EA / CB2 primer set (5 pmol / μl), 25 mM
The dNTP mix (TOYOBO) was thawed on ice, vortexed well, and the solution was recovered by Flash centrifugation. Using the reagents described above, 500 μl each of α and β (10 times) of a 2nd PCR solution was prepared on ice according to the following composition.
【００１９】表５ α、β鎖用２ndＰＣＲ溶液 Table 5 2nd PCR solution for α and β chains
【表５】 混合物を良く攪拌し、Flash遠心したのち、α、β各４
５０μｌ(10回分）の2ndＰＣＲ溶液を、先にｃＤＮＡを
分注しておいたチューブに４５μlずつ分注した。残っ
たＰＣＲ溶液のうち、18μlをネガティブコントロール
増幅に用いた。α、β各10本ずつのチューブにミネラル
オイル（SIGMA）を分注し、以下の手順でＰＣＲ増幅を
行った（変成94℃30秒を１サイクル、変成94℃１分、ア
ニール55℃、1.5分、伸長72℃、2分を２０サイクル、La
st extension72℃、5分以下で4℃で保存）。Flash遠心
後、-70℃に靜置し、氷結させた。室温に戻し、ミネラ
ルオイルのみが解凍した時点で、マイクロピペットによ
りこれを除去した。第2回のＰＣＲ産物５μｌに２μｌ
の電気泳動用色素を加えた。x１ＴＡＥ中で定電圧100ｍ
ｖの条件で、１％アガロース電気泳動を行い、サイズ分
画されたＤＮＡをエチジウムブロマイド染色し、トラン
スイルミネーターを用い、増幅を確認した。[Table 5] Stir the mixture well and centrifuge in Flash, then 4 each for α and β
50 μl (10 times) of the 2nd PCR solution was dispensed in 45 μl portions into the tube into which cDNA was previously dispensed. Of the remaining PCR solution, 18 μl was used for negative control amplification. Mineral oil (SIGMA) was dispensed into 10 tubes each of α and β, and PCR amplification was performed according to the following procedure (1 cycle of denaturation at 94 ° C for 30 seconds, denaturation at 94 ° C for 1 minute, annealing at 55 ° C and 1.5 Min, extension 72 ° C, 20 cycles of 2 min, La
st extension 72 ℃, stored at 4 ℃ for 5 minutes or less). After flash centrifugation, the mixture was allowed to stand at -70 ° C and frozen. After returning to room temperature and only the mineral oil was thawed, it was removed with a micropipette. 2 μl for 5 μl of second PCR product
Was added. Constant voltage 100m in x1 TAE
Under the conditions of v, 1% agarose electrophoresis was performed, the size-fractionated DNA was stained with ethidium bromide, and amplification was confirmed using a transilluminator.
【００２０】３）第３回（３rd）ＰＣＲ（ビオチン付
加） 上記２）で増幅を確認したＰＣＲ産物を、0.5ml滅菌チ
ューブに１μlずつ分注し氷上で保管した。x10 ＰＣＲ
用バッファー(Promega)、25mM MgCl2(Promega)、P20EA/
CA3及びP20EA/CB３プライマーセット（50pmol/μl）、1
0mM dATP, dCTP, dGTP mix, 0.5mMdTTP mix, 0.5mM Bio
tin-21-dUTPを氷上で解凍し、ボルテックスで良く攪拌
した後、Flash遠心により溶液を回収した。上記の試薬
を用い、α、β各1000μl（10回分）の３rdＰＣＲ溶液
を氷上で以下の組成に従い調製した。3) Third (3rd) PCR (addition of biotin) The PCR product confirmed to be amplified in the above 2) was dispensed at 1 μl per 0.5 ml sterile tube and stored on ice. x10 PCR
Buffer (Promega), 25 mM MgCl 2 (Promega), P20EA /
CA3 and P20EA / CB3 primer set (50 pmol / μl), 1
0mM dATP, dCTP, dGTP mix, 0.5mM dTTP mix, 0.5mM Bio
Tin-21-dUTP was thawed on ice, vortexed well, and the solution was recovered by Flash centrifugation. Using the reagents described above, 1000 μl (10 times) of 3rd PCR solution for each of α and β was prepared on ice according to the following composition.
【００２１】表６ α、β鎖用３rdＰＣＲ溶液 Table 6 3rd PCR solution for α and β chains
【表６】 混合物を良く攪拌し、Flash遠心したのち、α、β各１
０００μｌ(10回分）の3rdＰＣＲ溶液を、先に2ndｃＤ
ＮＡを分注しておいたチューブに１００μlずつ分注し
た。残ったＰＣＲR溶液のうち、２０μlをネガティブコ
ントロール増幅に用いた。α、β各10本ずつのチューブ
にミネラルオイル（SIGMA）を分注し、以下の手順でＰ
ＣＲ増幅を行った（変成94℃30秒を１サイクル、変成94
℃１分、アニール55℃、２分、伸長72℃、2.5分を２０
サイクル、Last extension72℃、5分以下で4℃で保
存）。Flash遠心後、-70℃に靜置し、氷結させた。室温
に戻し、ミネラルオイルのみが解凍した時点で、マイク
ロピペットによりこれを除去した。第３回のＰＣＲ産物
５μｌに２μｌの電気泳動用色素を加えた。x１ＴＡＥ
中で定電圧100ｍｖの条件で、１％アガロース電気泳動
を行い、サイズ分画されたＤＮＡをエチジウムブロマイ
ド染色し、トランスイルミネーターを用い、増幅を確認
した。[Table 6] Stir the mixture well, flash centrifuge, then each of α and β
2,000 μl (10 times) of the 3rd PCR solution was added to the 2ndcD
100 μl was dispensed into tubes into which NA had been dispensed. Of the remaining PCRR solution, 20 μl was used for negative control amplification. Dispense mineral oil (SIGMA) into 10 tubes each of α and β and follow the procedure below.
CR amplification was performed (1 cycle of denaturation 94 ° C for 30 seconds, denaturation 94
℃ 1 minute, annealing 55 ℃, 2 minutes, extension 72 ℃, 2.5 minutes 20
Cycle, Last extension 72 ° C, stored at 4 ° C for less than 5 minutes). After flash centrifugation, the mixture was allowed to stand at -70 ° C and frozen. After returning to room temperature and only the mineral oil was thawed, it was removed with a micropipette. To 5 μl of the third PCR product, 2 μl of electrophoresis dye was added. x1 TAE
In the medium, 1% agarose electrophoresis was performed under the condition of a constant voltage of 100 mv, and the size-fractionated DNA was stained with ethidium bromide, and amplification was confirmed using a transilluminator.
【００２２】ＩＩ． Reverse dot blotting Reverse dot blottingは、以下の表７〜１３に記載の、
各ＴＣＲＶサブファミリーに特異的に対応する２０bp前
後のオリゴヌクレオチドプローブを用い、常法に従って
行った。表７〜９には、ＴＣＲＶα及びＣαのための一
本鎖オリゴヌクレオチドプローブ（ＳＳＯＰ）、表１０
から１３には、ＴＣＲＶβ及びＣβのための一本鎖オリ
ゴヌクレオチドプローブ（ＳＳＯＰ）が記載されてい
る。表７〜９ Reverse dot blotting用ＴＣＲＶα及び
ＣαのためのＳＳＯＰII. Reverse dot blotting Reverse dot blotting is described in Tables 7 to 13 below.
This was carried out according to a conventional method using an oligonucleotide probe of about 20 bp specifically corresponding to each TCRV subfamily. Tables 7-9 show single stranded oligonucleotide probes (SSOP) for TCRVα and Cα, Table 10
To 13 describe single-stranded oligonucleotide probes (SSOPs) for TCRVβ and Cβ. Tables 7-9 SSOPs for TCRVα and Cα for Reverse dot blotting
【表７】 [Table 7]
【表８】 [Table 8]
【表９】 [Table 9]
【００２３】表１０〜１３ Reverse dot blotting用Ｔ
ＣＲＶβ及びＣβのためのＳＳＯＰ Tables 10-13 T for reverse dot blotting
SSOP for CRVβ and Cβ
【表１０】 [Table 10]
【表１１】 [Table 11]
【表１２】 [Table 12]
【表１３】 [Table 13]
【００２４】表７〜１3に記載のオリゴヌクレオチドを
合成し、poly-T-tailを付加し、標識し、供給者の指示
に従ってHybond Nfp メンブレン(アマーシャム社)に固
相化した。次いで、上記Ｉ.に記載のアダプター付加Ｐ
ＣＲで増幅した遺伝子群を、固相化したメンブレン上の
オリゴマーに３７℃で１時間、指定のバッファー中でハ
イブリダイズさせ、標識に用いたビオチンと、アルカリ
フォスファターゼ標識したアヴィジンによるＡＭＰＤＤ
の発光量の強度に基づいて検出することにより、同定、
定量した。なお、ＴＣＲＶβ２.１及びＶβ６.５に特異
的なオリゴプローブＶＢ０２−１及びＶＢ０６−４の塩
基配列は配列番号１０及び１１に示されている。Oligonucleotides shown in Tables 7 to 13 were synthesized, added with poly-T-tail, labeled, and immobilized on Hybond Nfp membrane (Amersham) according to the supplier's instructions. Next, the adapter-added P described in I. above.
The genes amplified by CR were hybridized with the oligomer on the immobilized membrane at 37 ° C. for 1 hour in a designated buffer, and the biotin used for labeling and the AMPDD with alkaline phosphatase-labeled avidin were used.
Identification by detecting based on the intensity of the luminescence amount of
Quantified. The nucleotide sequences of oligo probes VB02-1 and VB06-4 specific to TCRVβ2.1 and Vβ6.5 are shown in SEQ ID NOS: 10 and 11, respectively.
【００２５】川崎病患者２２名の治療前（発症期)、γ-
グロブリン投与時及び回復期における測定結果を図１及
び２に示す。さらに、川崎病患者２２名の治療前（発症
期）と回復期、発熱を伴う川崎病以外の疾患群の子供１
４名、及び健常大人１０名についてＴＣＲＶβ鎖のＶβ
２.１及びＶβ６.５の発現頻度を図３に示す。図１及び
２において、横軸のＶＡ及びＶＢは、表７〜１3に記載
の、Reverse dot blottingに用いたオリゴプローブの番
号を表し、縦軸は各サブファミリーの発現率（％）を示
す。なお、図１における１Ｄ，２Ｄ，３Ｄは、それぞ
れ、表２におけるＶＤ１，ＶＤ２，ＶＤ３に相当し、Ｖ
Ｂ０２−１はＶβ２.１を、ＶＢ０６−４はＶβ６.５を
表す。図１および２から、川崎病発症期にはＶβ鎖のＶ
β２.１及びＶβ６.５が高頻度で発現していることが分
かる また、図3からＴＣＲＶβ鎖については、川崎病発症期
の患者の場合、健常人コントロール及び川崎病回復期の
患者に比べて、Ｖβ２.１（P<0.05）及びＶβ６.５（P<
0.05）の使用頻度が有意に高いこと、他の疾患群と比較
した場合、Ｖβ６.５（P<0.01）の使用頻度が有意に高
いことが分かる。以上の結果は、川崎病の発症に関与す
る因子は、Ｖβ６.５陽性Ｔ細胞、又はＶβ６.５陽性及
びＶβ２.１陽性Ｔ細胞の増加をもたらすこと、即ち、
これらＴ細胞の増加に基づいて検出することが可能であ
ることを示している。Before treatment (onset) of 22 Kawasaki disease patients, γ-
The measurement results at the time of globulin administration and during the recovery period are shown in FIGS. In addition, 22 children with Kawasaki disease before treatment (onset period), convalescent period, and children with disease other than Kawasaki disease with fever.
VCR of TCRVβ chain for 4 and 10 healthy adults
The expression frequencies of 2.1 and Vβ6.5 are shown in FIG. In FIGS. 1 and 2, VA and VB on the horizontal axis represent the numbers of the oligo probes used for Reverse dot blotting described in Tables 7 to 13, and the vertical axis represents the expression rate (%) of each subfamily. 1D, 2D, and 3D in FIG. 1 correspond to VD1, VD2, and VD3 in Table 2, respectively.
B02-1 represents Vβ2.1 and VB06-4 represents Vβ6.5. From FIGS. 1 and 2, the Vβ chain V
It can be seen that β2.1 and Vβ6.5 are expressed at a high frequency. From FIG. 3, the TCRVβ chain is more evident in patients with Kawasaki disease onset than in healthy control and patients with Kawasaki disease recovery. , Vβ2.1 (P <0.05) and Vβ6.5 (P <0.05)
0.05) is significantly higher, and when compared to other disease groups, Vβ6.5 (P <0.01) is significantly higher. The above results indicate that the factor involved in the development of Kawasaki disease is an increase in Vβ6.5-positive T cells or Vβ6.5-positive and Vβ2.1-positive T cells,
This indicates that detection can be performed based on the increase in these T cells.
【００２６】ＩＩＩ．ＣＤＲ３領域の塩基配列の決定 治療前の発症期において、治療後の回復期より特に高い
割合でＶβ２.１及びＶβ６.５が発現しているサンプル
について、ＣＤＲ３領域（可変領域）の塩基配列を決定
した。即ち、アダプター付加ＰＣＲで増幅した産物を、
それぞれ、Ｖβ２.１及びＶβ６.５のための、ＶＢ０２
−１（表１０）とＣＢ３（表２）若しくはＶＢ０６−４
（表１１）とＣＢ３（表２）で、アダプター付加ＰＣＲ
と同じ条件下でＰＣＲ増幅し、アガロース電気泳動を行
った。ゲルから常法通り目的のＤＮＡを抽出した後、Ｔ
Ａクローニング法（Holton, T.A. and Ghanem, M.W. (1
991) A simple and efficient method for direct clon
ing of PCR products using ddT-tailed vectors Nucle
ic Acids Res 19, 1156)に従い、Ｖβ２.１及びＶβ６.
５の配列をもつＰＣＲ産物を、既知のベクター、ｐＧＥ
ＭＴのクローニングサイトに導入した。遺伝子導入され
たベクターを、電気穿孔法により、ＤＨ１０Ｂ由来の
Ｅ.coli K-12株に導入し、培養した。それぞれの形質転
換された大腸菌より抽出されたプラスミドは、Ｔ７ＤＮ
Ａポリメラーゼと、ＶＢ０２−１若しくはＶＢ０６−４
とＣＢ３プライマーのセットを用い、サンガー法(Sange
r ら、1987)によって塩基配列を決定した。該塩基配列
から予想されるアミノ酸配列を表１４及び表１５、１６
に示す。III. Determination of Nucleotide Sequence of CDR3 Region The nucleotide sequence of the CDR3 region (variable region) was determined for a sample in which Vβ2.1 and Vβ6.5 were expressed at a particularly high ratio during the onset period before treatment and during the recovery period after treatment. did. That is, the product amplified by the adapter-added PCR is
VB02 for Vβ2.1 and Vβ6.5, respectively.
-1 (Table 10) and CB3 (Table 2) or VB06-4
(Table 11) and CB3 (Table 2)
PCR was performed under the same conditions as described above, and agarose electrophoresis was performed. After extracting the target DNA from the gel as usual,
A cloning method (Holton, TA and Ghanem, MW (1
991) A simple and efficient method for direct clon
ing of PCR products using ddT-tailed vectors Nucle
ic Acids Res 19, 1156), Vβ2.1 and Vβ6.
The PCR product having a sequence of 5
It was introduced into the cloning site of MT. The transfected vector was introduced into DH10B-derived E. coli K-12 strain by electroporation and cultured. Plasmids extracted from each transformed E. coli were T7DN
A polymerase and VB02-1 or VB06-4
And Sanger method (Sange method)
r et al., 1987). Table 14 and Tables 15 and 16 show the amino acid sequence deduced from the nucleotide sequence.
Shown in
【００２７】表１４ 川崎病発症期の患者から得たＶβ
２.１−陽性ｃＤＮＡクローンのＣＤＲ３のアミノ酸配
列（ｎ＝２、クローン数１０又は１１） Table 14 Vβ obtained from patients at the onset of Kawasaki disease
2.1-Amino acid sequence of CDR3 of positive cDNA clone (n = 2, number of clones 10 or 11)
【表１４】 表１４に記載の２１クローンのアミノ酸配列は、上から
順次、配列番号９７〜配列番号１１７のアミノ酸配列
に、それぞれ対応している。[Table 14] The amino acid sequences of the 21 clones shown in Table 14 correspond to the amino acid sequences of SEQ ID NO: 97 to SEQ ID NO: 117, respectively, from the top.
【００２８】表１５〜１６ 川崎病発症期の患者から得
たＶβ６.５−陽性ｃＤＮＡクローンのＣＤＲ３のアミ
ノ酸配列（ｎ＝４、クローン数１０〜１２） Table 15-16 Amino acid sequence of CDR3 of Vβ6.5-positive cDNA clone obtained from Kawasaki disease onset patient (n = 4, number of clones 10-12)
【表１５】 [Table 15]
【表１６】 表１５に記載の２０クローンのアミノ酸配列は、配列番
号１１８〜１３７、表１６に記載の２３クローンのアミ
ノ酸配列は、配列番号１３８〜１６０に記載のアミノ酸
配列に、それぞれ対応している。表１４〜１６から明ら
かなように、同一のクローンは存在していない。これ
は、川崎病発症期におけるＶβ２.１、Ｖβ６.５ポジテ
ィブＴ細胞の発現量増加が、ポリクローナルであること
を示している。[Table 16] The amino acid sequences of the 20 clones shown in Table 15 correspond to SEQ ID NOs: 118 to 137, and the amino acid sequences of the 23 clones shown in Table 16 correspond to the amino acid sequences shown in SEQ ID NOs: 138 to 160, respectively. As is clear from Tables 14 to 16, no identical clone exists. This indicates that the increase in the expression level of Vβ2.1 and Vβ6.5 positive T cells in the onset stage of Kawasaki disease is polyclonal.
【００２９】ＩＶ．各種サイトカインの測定 川崎病（発症期及び回復期）及び他の疾患群の血清につ
いて、Quantikine immunoassay kit (R & D system
社)、Cytoscreen immunoassay kit (BioSouce Internat
ional 社)、Human Interferon Gamma ELISA test kit(B
RL社)を用い、ＩＬ−１β、ＩＬ−２、ＩＬ−６、ＩＬ
−８、ＩＬ−１０、ＩＦＮγ、Ｇ−ＣＳＦ、ＴＮＦ−α
量を測定した。結果を表１7に示す。表１7 川崎病の発症期及び回復期、及び他の疾患の患
者の、血清サイトカイン濃度IV. Measurement of various cytokines Serum from Kawasaki disease (onset and recovery) and other disease groups was analyzed using the Quantikine immunoassay kit (R & D system).
), Cytoscreen immunoassay kit (BioSouce Internat
ional), Human Interferon Gamma ELISA test kit (B
RL), IL-1β, IL-2, IL-6, IL
-8, IL-10, IFNγ, G-CSF, TNF-α
The amount was measured. The results are shown in Table 17. Table 17 Serum cytokine levels in patients with Kawasaki disease onset and recovery, and other diseases
【表１７】 注：ａ）検出可能なサイトカイン濃度以下の血清濃度を
ゼロとして、平均値を算出した。表から、各種炎症性の
サイトカイン産生もＴＣＲ Ｖβ2.1やＶβ６.５同様、
川崎病回復期に比べ発症期において高い傾向が見られ
る。また、他の疾患群と比較した場合も、多くのサイト
カインが、その発症期で高い傾向にあることが分かる。
この結果は、末梢血中でＴ細胞が増加していることを示
している。以上の結果は、川崎病の発症には、スーパー
抗原が関与していること、さらには、Ａ群連鎖球菌由来
の外毒素 (SPEC)の関与を示唆するものである。[Table 17] Notes: a) The mean value was calculated with the serum concentration below the detectable cytokine concentration as zero. From the table, the production of various inflammatory cytokines was similar to that of TCR Vβ2.1 and Vβ6.5.
A higher tendency is seen in the onset period than in the Kawasaki disease recovery period. Also, when compared with other disease groups, it can be seen that many cytokines tend to be high in the onset period.
This result indicates that T cells are increased in peripheral blood. The above results suggest that superantigens are involved in the development of Kawasaki disease, and that exotoxins (SPEC) derived from group A streptococci are involved.
【００３０】[0030]
【産業上の有用性】原因が未解明であり診断の確定に時
間を要するために、重篤な症状に至る例が多い川崎病に
関連する因子を、その発症期に科学的な方法で検出する
手段を提供することにより、川崎病の早期診断、早期治
療を可能にすることができる。[Industrial usefulness] Factors related to Kawasaki disease that often lead to serious symptoms are detected by scientific methods during the onset period because the cause is unknown and it takes time to confirm the diagnosis. By providing means for performing the above, early diagnosis and early treatment of Kawasaki disease can be made possible.
【００３１】[0031]
【配列表】 <110> 塩野義製薬株式会社 Shionogi & Co., Ltd. <120> 川崎病因子の検出法 <130> 155380 <160> 160 <210> 1 <211> 10 <212> DNA <213> Artificial Sequence <400> 1 gggaattcgg 10 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <400> 2 taatacgact ccgaattccc 20 <210> 3 <211> 45 <212> DNA <213> Artificial Sequence <400> 3 gactagtcaa agcggccgcg agctcttttt tttttttttt ttttt 45 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <400> tgttgaaggc gtttgcacat gca 23 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <400> 5 gtccatagac ctcatgtcta gca 3 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <400> 6 actttgtgac acatttgttt gag 23 <210> 7 <211> 23 <212> DNA <213> Artificial Sequence <400> 7 gaactggact tgacagcgga agt 23 <210> 8 <211> 23 <212> DNA <213> Artificial Sequence <400> 8 aggcagtatc tggagtcatt gag 23 <210> 9 <211> 23 <212> DNA <213> Artificial Sequence <400> 9 actgtgcacc tccttcccat tca 23 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <400> 10 tcatcaacca tgcaagcctg a 21 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <400> 11 aaggctgctc agtgatcggt 20 <210> 12 <211> 23 <212> DNA <213> Artificial Sequence <400> 12 gaaacaagtt ggtggtcata tta 23 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <400> 13 aaaggagaag cgatcggtaa c 21 <210> 14 <211> 23 <212> DNA <213> Artificial Sequence <400> 14 gacactgtat attcaaatcc aga 23 <210> 15 <211> 21 <212> DNA <213> Artificial Sequence <400> 15 atcaagggct ttgaggctga a 21 <210> 16 <211> 22 <212> DNA <213> Artificial Sequence <400> 16 atcaacggtt ttgaggctga at 22 <210> 17 <211> 21 <212> DNA <213> Artificial Sequence <400> 17 gcattaaagg ctttgaggct g 21 <210> 18 <211> 21 <212> DNA <213> Artificial Sequence <400> 18 catcacggtt ttgaggctga a 21 <210> 19 <211> 21 <212> DNA <213> Artificial Sequence <400> 19 acagctcaat aaagccagcc a 21 <210> 20 <211> 21 <212> DNA <213> Artificial Sequence <400> 20 cacaggtcga taaatccagc a 21 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <400> 21 agagccagcc agtatatttc c 21 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <400> 22 gtggaagatt aagagtcacg c 21 <210> 23 <211> 21 <212> DNA <213> Artificial Sequence <400> 23 aacaacagaa tggcctctct g 21 <210> 24 <211> 21 <212> DNA <213> Artificial Sequence <400> 24 cctctctgat catcacagaa g 21 <210> 25 <211> 21 <212> DNA <213> Artificial Sequence <400> 25 ctgaaggtca cctttgatac c 21 <210> 26 <211> 21 <212> DNA <213> Artificial Sequence <400> 26 tgacaccagt gatccaagtt a 21 <210> 27 <211> 22 <212> DNA <213> Artificial Sequence <400> 27 attccttagt cgctctgata gt 22 <210> 28 <211> 21 <212> DNA <213> Artificial Sequence <400> 28 ttccttagtc ggtctaaagg g 21 <210> 29 <211> 21 <212> DNA <213> Artificial Sequence <400> 29 tgggcgaaaa gaaagaccaa c 21 <210> 30 <211> 21 <212> DNA <213> Artificial Sequence <400> 30 ggccaaagag tcaccgtttt a 21 <210> 31 <211> 21 <212> DNA <213> Artificial Sequence <400> 31 catcaaaggc ttcactgctg a 21 <210> 32 <211> 21 <212> DNA <213> Artificial Sequence <400> 32 gtggagaagt gaagaagctg a 21 <210> 33 <211> 21 <212> DNA <213> Artificial Sequence <400> 33 agggacgata caacatgacc t 21 <210> 34 <211> 21 <212> DNA <213> Artificial Sequence <400> 34 gtcggtattc ttggaacttc c 21 <210> 35 <211> 21 <212> DNA <213> Artificial Sequence <400> 35 aagattaagc gccacgactg t 21 <210> 36 <211> 21 <212> DNA <213> Artificial Sequence <400> 36 cgtttctctg tgaacttcca g 21 <210> 37 <211> 21 <212> DNA <213> Artificial Sequence <400> 37 gaccaaagac tcactgttct a 21 <210> 38 <211> 21 <212> DNA <213> Artificial Sequence <400> 38 cagctatggc tttgaagctg a 21 <210> 39 <211> 22 <212> DNA <213> Artificial Sequence <400> 39 gaaggaagat tcacaatctc ct 22 <210> 40 <211> 21 <212> DNA <213> Artificial Sequence <400> 40 ggacgatata gtgccactct t 21 <210> 41 <211> 22 <212> DNA <213> Artificial Sequence <400> 41 gcatggaaga ttaattgcca ca 22 <210> 42 <211> 23 <212> DNA <213> Artificial Sequence <400> 42 gactatacta acagcatgtt tga 23 <210> 43 <211> 21<212> DNA <213> Artificial Sequence <400> 43 aacaaaagtg ccaagcacct c 21 <210> 44 <211> 21 <212> DNA <213> Artificial Sequence <400> 44 tgacaaggga agcaacaaag g 21 <210> 45 <211> 21 <212> DNA <213> Artificial Sequence <400> 45 aagtggaaga cttaatgccc c 21 <210> 46 <211> 21 <212> DNA <213> Artificial Sequence <400> 46 caactctgga tgcagacaca a 21 <210> 47 <211> 21 <212> DNA <213> Artificial Sequence <400> 47 aagactgact gctcagtttg g 21 <210> 48 <211> 22 <212> DNA <213> Artificial Sequence <400> 48 agataactgc caagttggat ga 22 <210> 49 <211> 21 <212> DNA <213> Artificial Sequence <400> 49 ggacgattaa tggcctcact t 21 <210> 50 <211> 22 <212> DNA <213> Artificial Sequence <400> 50 gtcaggaaga ctaagtagca ta 22 <210> 51 <211> 20 <212> DNA <213> Artificial Sequence <400> 51 ggagaacaga tgcgtcgtga 20 <210> 52 <211> 21 <212> DNA <213> Artificial Sequence <400> 52 tggagaacag aagggtcatg a 21 <210> 53 <211> 22 <212> DNA <213> Artificial Sequence <400> 53 tgaagaagca gaaaagactg ac 22 <210> 54 <211> 21 <212> DNA <213> Artificial Sequence <400> 54 ctgtattcag ctggggaaga a 21 <210> 55 <211> 22 <212> DNA <213> Artificial Sequence <400> 55 tcagagagag acaatggaaa ac 22 <210> 56 <211> 20 <212> DNA <213> Artificial Sequence <400> 56 tatccagaac cctgaccctg 20 <210> 57 <211> 21 <212> DNA <213> Artificial Sequence <400> 57 tgtaccagct gagagactct a 21 <210> 58 <211> 21 <212> DNA <213> Artificial Sequence <400> 58 acaacagttc cctgacttgc a 21 <210> 59 <211> 21 <212> DNA <213> Artificial Sequence <400> 59 ccatcaacca tccaaacctg a 21 <210> 60 <211> 21 <212> DNA <213> Artificial Sequence <400> 60 gtctctagag agaagaagga g 21 <210> 61 <211> 21 <212> DNA <213> Artificial Sequence <400> 61 ttgacaagtt tcccatcagc c 21 <210> 62 <211> 21 <212> DNA <213> Artificial Sequence <400> 62 cttcagtgag acacagagaa a 21 <210> 63 <211> 22 <212> DNA <213> Artificial Sequence <400> 63 attatgagga ggaagagaga ca 22 <210> 64 <211> 23 <212> DNA <213> Artificial Sequence <400> 64 tattatgaga aagaagagag agg 23 <210> 65 <211> 21 <212> DNA <213> Artificial Sequence <400> 65 tgacgagggt gaagagagaa a 21 <210> 66 <211> 22 <212> DNA <213> Artificial Sequence <400> 66 attataggga ggaagagaat gg 22 <210> 67 <211> 21 <212> DNA <213> Artificial Sequence <400> 67 acgatcggtt ctttgcagtc a 21 <210> 68 <211> 21 <212> DNA <213> Artificial Sequence <400> 68 tcaactagac aaatcggggc t 21 <210> 69 <211> 21 <212> DNA <213> Artificial Sequence <400> 69 ccaacaagac aaatcagggc t 21 <210> 70 <211> 20 <212> DNA <213> Artificial Sequence <400> 70 tcgcttctct gcagagagga 20 <210> 71 <211> 21 <212> DNA <213> Artificial Sequence <400> 71 aactggaaaa atcagggctg c 21 <210> 72 <211> 20<212> DNA <213> Artificial Sequence <400> 72 ctgaatgccc caacagctct 20 <210> 73 <211> 23 <212> DNA <213> Artificial Sequence <400> 73 tttactttaa caacaacgtt ccg 23 <210> 74 <211> 20 <212> DNA <213> Artificial Sequence <400> 74 aaggactgga gttgctggct 20 <210> 75 <211> 22 <212> DNA <213> Artificial Sequence <400> 75 aaacagttcc aaatcgcttc tc 22 <210> 76 <211> 22 <212> DNA <213> Artificial Sequence <400> 76 ccaaaaactc atcctgtacc tt 22 <210> 77 <211> 22 <212> DNA <213> Artificial Sequence <400> 77 tcaacagtct ccagaataag ga 22 <210> 78 <211> 21 <212> DNA <213> Artificial Sequence <400> 78 caaaggagaa gtctcagatg g 21 <210> 79 <211> 21 <212> DNA <213> Artificial Sequence <400> 79 caatggctac aatgtctcca g 21 <210> 80 <211> 20 <212> DNA <213> Artificial Sequence <400> 80 ggtccctgat ggctacaatg 20 <210> 81 <211> 21 <212> DNA <213> Artificial Sequence <400> 81 tgatggttat agtgtctcca g 21 <210> 82 <211> 20 <212> DNA <213> Artificial Sequence <400> 82 ccgaatggct acaacgtctc 20 <210> 83 <211> 21 <212> DNA <213> Artificial Sequence <400> 83 gtctctcgaa aagagaagag g 21 <210> 84 <211> 20 <212> DNA <213> Artificial Sequence <400> 84 agtgtctctc gacaggcaca 20 <210> 85 <211> 21 <212> DNA <213> Artificial Sequence <400> 85 gtgaaagagt ctaaacagga t 21 <210> 86 <211> 23 <212> DNA <213> Artificial Sequence <400> 86 cacagatagt aaatgacttt cag 23 <210> 87 <211> 21 <212> DNA <213> Artificial Sequence <400> 87 gatgagtcag gaatgccaaa g 21 <210> 88 <211> 20 <212> DNA <213> Artificial Sequence <400> 88 ctcaatgccc caagaacgca 20 <210> 89 <211> 20 <212> DNA <213> Artificial Sequence <400> 89 tctgaggtgc cccagaatct 20 <210> 90 <211> 20 <212> DNA <213> Artificial Sequence <400> 90 tctgcagaga ggctcaaagg 20 <210> 91 <211> 23 <212> DNA <213> Artificial Sequence <400> 91 ttcagtgact atcattctga act 23 <210> 92 <211> 21 <212> DNA <213> Artificial Sequence <400> 92 attcttgggg cagaaagtcg a 21 <210> 93 <211> 19 <212> DNA <213> Artificial Sequence <400> 93 ccaggaggcc gaacacttc 19 <210> 94 <211> 21 <212> DNA <213> Artificial Sequence <400> 94 aacaggtatg cccaaggaaa g 21 <210> 95 <211> 19 <212> DNA <213> Artificial Sequence <400> 95 cacccgaggt cgctgtgtt 19 <210> 96 <211> 21 <212> DNA <213> Artificial Sequence <400> 96 gctgtgtttg agccatcaga a 21 <210> 97 <211> 27 <212> PRT <213> Homo sapiens <400> 97 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Val Gly Asn Gly Tyr 1 5 10 15 Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 <210> 98 <211> 30 <212> PRT <213> Homo sapiens <400> 98 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Asp Glu Ala Ile 1 5 10 15 Thr Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 99 <211> 31 <212> PRT <213> Homo sapiens <400> 99 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Asp Pro Thr Gly Arg 1 5 10 15 His Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 100 <211> 29 <212> PRT <213> Homo sapiens <400> 100 Glu Asp Ser Ser Phe Tyr Ile Cys Arg Pro Ser Ser Gly Ser Tyr Asn 1 5 10 15 Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 101 <211> 34 <212> PRT <213> Homo sapiens <400> 101 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Leu Ala Gly Thr 1 5 10 15 Ser Ser Ser Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr 20 25 30 Val Leu <210> 102 <211> 34 <212> PRT <213> Homo sapiens <400> 102 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ile Gly Thr Thr Ser Gly 1 5 10 15 Ser His Arg Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr 20 25 30 Val Leu <210> 103 <211> 26 <212> PRT <213> Homo sapiens <400> 103 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ser Gly Glu Thr Gln Tyr 1 5 10 15 Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 104 <211> 30 <212> PRT <213> Homo sapiens <400> 104 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Pro Ser Gly Arg Ala Gln 1 5 10 15 Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 105 <211> 28 <212> PRT <213> Homo sapiens <400> 105 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Val Gly Gly Gln Ser Tyr Glu 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 106 <211> 28 <212> PRT <213> Homo sapiens <400> 106 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ala Gln Asp Val Tyr Glu 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 107 <211> 29 <212> PRT <213> Homo sapiens <400> 107 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Asp Thr Arg Gly Met Asn Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 108 <211> 31 <212> PRT <213> Homo sapiens <400> 108 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Ala Asp Arg Tyr 1 5 10 15 Asn Thr Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 109 <211> 29 <212> PRT <213> Homo sapiens <400> 109 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Glu Thr Ser Asn Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 110 <211> 28 <212> PRT <213> Homo sapiens <400> 110 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Phe Ser Ser Gly Gly 1 5 10 15 Tyr Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 <210> 111 <211> 30 <212> PRT <213> Homo sapiens <400> 111 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Asn Leu Ala Gly Val Gly 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 112 <211> 30 <212> PRT <213> Homo sapiens <400> 112 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Val Ser Gly Arg Thr 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 113 <211> 31 <212> PRT <213> Homo sapiens <400> 113 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Tyr Thr Ser Gly Arg Ser 1 5 10 15 Lys Asn Ile Gln Tyr Phe Gly Ala Gly Thr Arg Leu Ser Val Leu 20 25 30 <210> 114 <211> 29 <212> PRT <213> Homo sapiens <400> 114 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Leu Ala Val Glu 1 5 10 15 Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 115 <211> 31 <212> PRT <213> Homo sapiens <400> 115 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Pro Ile Val Gly Gly 1 5 10 15 Ser His Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 116 <211> 30 <212> PRT <213> Homo sapiens <400> 116 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Pro Gly Thr Ser 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 117 <211> 30 <212> PRT <213> Homo sapiens <400> 117 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Thr Ser Ser Gly Ser Ser 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 118 <211> 29 <212> PRT <213> Homo sapiens <400> 118 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Glu Asp Val Gly Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 119 <211> 32 <212> PRT <213> Homo sapiens <400> 119 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Thr Ser Gly Gly 1 5 10 15 Ala Trp Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 120 <211> 29 <212> PRT <213> Homo sapiens <400> 120 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ile Lys Thr Asp Pro Arg Arg 1 5 10 15 Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 121 <211> 28 <212> PRT <213> Homo sapiens <400> 121 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Arg Ser Gly Gly Gly Glu 1 5 10 15 Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 <210> 122 <211> 33 <212> PRT <213> Homo sapiens <400> 122 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gln Gly 1 5 10 15 Gln Gly Lys Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Leu <210> 123 <211> 31 <212> PRT <213> Homo sapiens <400> 123 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Arg Leu Ala Gly Ala Phe 1 5 10 15 Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 124 <211> 28 <212> PRT <213> Homo sapiens <400> 124 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Ile Thr Asp Thr 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 125 <211> 32 <212> PRT <213> Homo sapiens <400> 125 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Gly Gly Gly 1 5 10 15 Arg Val Ala Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 126 <211> 31 <212> PRT <213> Homo sapiens <400> 126 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Gly Gln Glu 1 5 10 15 Asn Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 127 <211> 32 <212> PRT <213> Homo sapiens <400> 127 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Arg Ala Ser Gly 1 5 10 15 Arg Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 128 <211> 27 <212> PRT <213> Homo sapiens <400> 128 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Glu Leu Gly Phe 1 5 10 15 Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 <210> 129 <211> 29 <212> PRT <213> Homo sapiens <400> 129 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Phe Gly Gly Glu 1 5 10 15 Lys Leu Phe Phe Gly Ser Gly Thr Gln Leu Ser Val Leu 20 25 <210> 130 <211> 29 <212> PRT <213> Homo sapiens <400> 130 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Arg Asp Gln 1 5 10 15 Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 <210> 131 <211> 30 <212> PRT <213> Homo sapiens <400> 131 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Pro Gln Ser Gly His Ala 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 132 <211> 28 <212> PRT <213> Homo sapiens <400> 132 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Phe Thr Asp Thr 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 133 <211> 26 <212> PRT <213> Homo sapiens <400> 133 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Pro Pro Gln Tyr 1 5 10 15 Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 134 <211> 27 <212> PRT <213> Homo sapiens <400> 134 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Lys Trp Thr Gln 1 5 10 15 Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 135 <211> 27 <212> PRT <213> Homo sapiens <400> 135 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Arg Glu Gly Gly Gln 1 5 10 15 Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 136 <211> 31 <212> PRT <213> Homo sapiens <400> 136 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Gly Ser Leu Ser 1 5 10 15 Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 137 <211> 30 <212> PRT <213> Homo sapiens <400> 137 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Ala Gln Gly Ala 1 5 10 15 Ala Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 138 <211> 30 <212> PRT <213> Homo sapiens <400> 138 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Gly Ser Leu Val Asp 1 5 10 15 Gln Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 30 <210> 139 <211> 30 <212> PRT <213> Homo sapiens <400> 139 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Gln Gln Gly Gly Gly 1 5 10 15 Asn Thr Ile Tyr Phe Gly Glu Gly Ser Trp Leu Thr Val Val 20 25 30 <210> 140 <211> 31 <212> PRT <213> Homo sapiens <400> 140 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Ser Leu Val 1 5 10 15 Asp Gln Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 30 <210> 141 <211> 30 <212> PRT <213> Homo sapiens <400> 141 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ile Gly Ala Ala 1 5 10 15 His Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 142 <211> 27 <212> PRT <213> Homo sapiens <400> 142 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Asp Glu Arg Glu 1 5 10 15 Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 143 <211> 31 <212> PRT <213> Homo sapiens <400> 143 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Ser Thr Arg 1 5 10 15 Gly Asp Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 144 <211> 31 <212> PRT <213> Homo sapiens <400> 144 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Gln Gly Gly Gly Gly 1 5 10 15 Thr Gly Glu Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 30 <210> 145 <211> 33 <212> PRT <213> Homo sapiens <400> 145 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Leu Val Arg 1 5 10 15 Asp Tyr Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Thr <210> 146 <211> 30 <212> PRT <213> Homo sapiens <400> 146 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Thr Gly Gly Gly 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 147 <211> 30 <212> PRT <213> Homo sapiens <400> 147 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Thr Thr Asp Ala 1 5 10 15 Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 148 <211> 31 <212> PRT <213> Homo sapiens <400> 148 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ala Ile Ala Gly Ala 1 5 10 15 Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 149 <211> 30 <212> PRT <213> Homo sapiens <400> 149 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ala Gly Gln Gly Ser 1 5 10 15 Thr Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 150 <211> 31 <212> PRT <213> Homo sapiens <400> 150 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Leu Val Gly Gly 1 5 10 15 Arg Gly Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 151 <211> 30 <212> PRT <213> Homo sapiens <400> 151 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Leu Gly Gln Asn 1 5 10 15 Tyr Gly Tyr Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 152 <211> 27 <212> PRT <213> Homo sapiens <400> 152 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Val Asp Thr Ile 1 5 10 15 Tyr Phe Gly Glu Gly Ser Trp Leu Thr Val Val 20 25 <210> 153 <211> 32 <212> PRT <213> Homo sapiens <400> 153 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Gly Thr Ser Gly 1 5 10 15 Ser Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 154 <211> 31 <212> PRT <213> Homo sapiens <400> 154 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Gly Arg Gly Asn 1 5 10 15 Thr Gly Glu Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 30 <210> 155 <211> 33 <212> PRT <213> Homo sapiens <400> 155 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Ile Arg Gly 1 5 10 15 Ala Ser Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Leu <210> 156 <211> 31 <212> PRT <213> Homo sapiens <400> 156 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Thr Ser Gly 1 5 10 15 Ala Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 157 <211> 31 <212> PRT <213> Homo sapiens <400> 157 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Val Thr Pro 1 5 10 15 Gln Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 158 <211> 36 <212> PRT <213> Homo sapiens <400> 158 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ile Pro Arg Leu Gly 1 5 10 15 Gln Gly Ala Ser Gly Ala Asn Val Leu Thr Phe Gly Ala Gly Ser Arg 20 25 30 Leu Thr Val Leu 35 <210> 159 <211> 30 <212> PRT <213> Homo sapiens <400> 159 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Thr Gly Gly 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 160 <211> 32 <212> PRT <213> Homo sapiens <400> 160 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Ile Pro Gly Leu 1 5 10 15 Arg Asn Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30[Sequence List] <110> Shionogi & Co., Ltd. <120> Detection method of Kawasaki disease factor <130> 155 380 <160> 160 <210> 1 <211> 10 <212> DNA <213 > Artificial Sequence <400> 1 gggaattcgg 10 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <400> 2 taatacgact ccgaattccc 20 <210> 3 <211> 45 <212> DNA <213> Artificial Sequence <400> 3 gactagtcaa agcggccgcg agctcttttt tttttttttt ttttt 45 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <400> tgttgaaggc gtttgcacat gca 23 <210> 5 <211> 23 <212> DNA <213> Artificial Sequence <400> 5 gtccatagac ctcatgtcta gca 3 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <400> 6 actttgtgac acatttgttt gag 23 <210> 7 <211> 23 <212> DNA <213> Artificial Sequence <400> 7 gaactggact tgacagcgga agt 23 <210> 8 <211> 23 <212> DNA <213> Artificial Sequence <400> 8 aggcagtatc tggagtcatt gag 23 <210> 9 <211> 23 <212> DNA <213> Artificial Sequence <400> 9 actgtgcacc tccttcccat tca 23 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <400> 10 tcatcaacca tgcaagcctg a 21 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <400> 11 aaggctgctc agtgatcggt 20 <210> 12 <211> 23 <212> DNA <213> Artificial Sequence <400> 12 gaaacaagtt ggtggtcata tta 23 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <400> 13 aaaggagaag cgatcggtaa c 21 <210> 14 <211> 23 <212> DNA <213> Artificial Sequence <400> 14 gacactgtat attcaaatcc aga 23 <210> 15 <211> 21 <212> DNA <213> Artificial Sequence <400> 15 atcaagggct ttgaggctga a 21 <210> 16 <211> 22 <212> DNA <213> Artificial Sequence <400> 16 atcaacggtt ttgaggctga at 22 <210> 17 <211> 21 <212> DNA <213> Artificial Sequence <400> 17 gcattaaagg ctttgaggct g 21 <210> 18 <211> 21 <212> DNA <213> Artificial Sequence <400> 18 catcacggtt ttgaggctga a 21 <210> 19 <211> 21 <212> DNA <213> Artificial Sequence <400> 19 acagctcaat aaagccagcc a 21 <210> 20 <211> 21 <212> DNA <213> Artificial Sequence <400> 20 cacaggtcga taaatccagc a 21 <210> 21 <211> 21 <212> DNA <213> Artificial Sequence <400> 21 agag ccagcc agtatatttc c 21 <210> 22 <211> 21 <212> DNA <213> Artificial Sequence <400> 22 gtggaagatt aagagtcacg c 21 <210> 23 <211> 21 <212> DNA <213> Artificial Sequence <400> 23 aacaacagaa tggcctctct g 21 <210> 24 <211> 21 <212> DNA <213> Artificial Sequence <400> 24 cctctctgat catcacagaa g 21 <210> 25 <211> 21 <212> DNA <213> Artificial Sequence <400> 25 ctgaaggtca cctttgatac c 21 <210> 26 <211> 21 <212> DNA <213> Artificial Sequence <400> 26 tgacaccagt gatccaagtt a 21 <210> 27 <211> 22 <212> DNA <213> Artificial Sequence <400> 27 attccttagt cgctctgata gt 22 <210> 28 <211> 21 <212> DNA <213> Artificial Sequence <400> 28 ttccttagtc ggtctaaagg g 21 <210> 29 <211> 21 <212> DNA <213> Artificial Sequence <400> 29 tgggcgaaaa gaaagaccaa c 21 <210> 30 <211> 21 <212> DNA <213> Artificial Sequence <400> 30 ggccaaagag tcaccgtttt a 21 <210> 31 <211> 21 <212> DNA <213> Artificial Sequence <400> 31 catcaaaggc ttcactgctg a 21 <210> 32 <211> 21 <212> DNA <213> Artificial Sequence <400> 32 gtggagaagt ga agaagctg a 21 <210> 33 <211> 21 <212> DNA <213> Artificial Sequence <400> 33 agggacgata caacatgacc t 21 <210> 34 <211> 21 <212> DNA <213> Artificial Sequence <400> 34 gtcggtattc ttggaacttc c 21 <210> 35 <211> 21 <212> DNA <213> Artificial Sequence <400> 35 aagattaagc gccacgactg t 21 <210> 36 <211> 21 <212> DNA <213> Artificial Sequence <400> 36 cgtttctctg tgaacttcca g 21 <210> 37 <211> 21 <212> DNA <213> Artificial Sequence <400> 37 gaccaaagac tcactgttct a 21 <210> 38 <211> 21 <212> DNA <213> Artificial Sequence <400> 38 cagctatggc tttgaagctg a 21 <210> 39 <211> 22 <212> DNA <213> Artificial Sequence <400> 39 gaaggaagat tcacaatctc ct 22 <210> 40 <211> 21 <212> DNA <213> Artificial Sequence <400> 40 ggacgatata gtgccactct t 21 <210> 41 <211> 22 <212> DNA <213> Artificial Sequence <400> 41 gcatggaaga ttaattgcca ca 22 <210> 42 <211> 23 <212> DNA <213> Artificial Sequence <400> 42 gactatacta acagcatgtt tga 23 <210> 43 <211> 21 <212> DNA <213> Artificial Sequence <400> 43 aacaaaagtg ccaagcacc tc 21 <210> 44 <211> 21 <212> DNA <213> Artificial Sequence <400> 44 tgacaaggga agcaacaaag g 21 <210> 45 <211> 21 <212> DNA <213> Artificial Sequence <400> 45 aagtggaaga cttaatgccc c 21 <210> 46 <211> 21 <212> DNA <213> Artificial Sequence <400> 46 caactctgga tgcagacaca a 21 <210> 47 <211> 21 <212> DNA <213> Artificial Sequence <400> 47 aagactgact gctcagtttg g 21 <210> 48 <211> 22 <212> DNA <213> Artificial Sequence <400> 48 agataactgc caagttggat ga 22 <210> 49 <211> 21 <212> DNA <213> Artificial Sequence <400> 49 ggacgattaa tggcctcact t 21 <210> 50 <211> 22 <212> DNA <213> Artificial Sequence <400> 50 gtcaggaaga ctaagtagca ta 22 <210> 51 <211> 20 <212> DNA <213> Artificial Sequence <400> 51 ggagaacaga tgcgtcgtga 20 <210> 52 <211> 21 <212> DNA <213> Artificial Sequence <400> 52 tggagaacag aagggtcatg a 21 <210> 53 <211> 22 <212> DNA <213> Artificial Sequence <400> 53 tgaagaagca gaaaagactg ac 22 <210> 54 <211> 21 <212> DNA <213> Artificial Sequence <400> 54 ctgtattcag ctggggaaga a 21 <2 10> 55 <211> 22 <212> DNA <213> Artificial Sequence <400> 55 tcagagagag acaatggaaa ac 22 <210> 56 <211> 20 <212> DNA <213> Artificial Sequence <400> 56 tatccagaac cctgaccctg 20 <210 > 57 <211> 21 <212> DNA <213> Artificial Sequence <400> 57 tgtaccagct gagagactct a 21 <210> 58 <211> 21 <212> DNA <213> Artificial Sequence <400> 58 acaacagttc cctgacttgc a 21 <210 > 59 <211> 21 <212> DNA <213> Artificial Sequence <400> 59 ccatcaacca tccaaacctg a 21 <210> 60 <211> 21 <212> DNA <213> Artificial Sequence <400> 60 gtctctagag agaagaagga g 21 <210 > 61 <211> 21 <212> DNA <213> Artificial Sequence <400> 61 ttgacaagtt tcccatcagc c 21 <210> 62 <211> 21 <212> DNA <213> Artificial Sequence <400> 62 cttcagtgag acacagagaa a 21 <210 > 63 <211> 22 <212> DNA <213> Artificial Sequence <400> 63 attatgagga ggaagagaga ca 22 <210> 64 <211> 23 <212> DNA <213> Artificial Sequence <400> 64 tattatgaga aagaagagag agg 23 <210 > 65 <211> 21 <212> DNA <213> Artificial Sequence <400> 65 tgacgagggt gaagagagaa a 21 <210> 66 < 211> 22 <212> DNA <213> Artificial Sequence <400> 66 attataggga ggaagagaat gg 22 <210> 67 <211> 21 <212> DNA <213> Artificial Sequence <400> 67 acgatcggtt ctttgcagtc a 21 <210> 68 < 211> 21 <212> DNA <213> Artificial Sequence <400> 68 tcaactagac aaatcggggc t 21 <210> 69 <211> 21 <212> DNA <213> Artificial Sequence <400> 69 ccaacaagac aaatcagggc t 21 <210> 70 < 211> 20 <212> DNA <213> Artificial Sequence <400> 70 tcgcttctct gcagagagga 20 <210> 71 <211> 21 <212> DNA <213> Artificial Sequence <400> 71 aactggaaaa atcagggctg c 21 <210> 72 <211 > 20 <212> DNA <213> Artificial Sequence <400> 72 ctgaatgccc caacagctct 20 <210> 73 <211> 23 <212> DNA <213> Artificial Sequence <400> 73 tttactttaa caacaacgtt ccg 23 <210> 74 <211> 20 <212> DNA <213> Artificial Sequence <400> 74 aaggactgga gttgctggct 20 <210> 75 <211> 22 <212> DNA <213> Artificial Sequence <400> 75 aaacagttcc aaatcgcttc tc 22 <210> 76 <211> 22 <212> DNA <213> Artificial Sequence <400> 76 ccaaaaactc atcctgtacc tt 22 <210> 77 <211> 22 <212 > DNA <213> Artificial Sequence <400> 77 tcaacagtct ccagaataag ga 22 <210> 78 <211> 21 <212> DNA <213> Artificial Sequence <400> 78 caaaggagaa gtctcagatg g 21 <210> 79 <211> 21 <212 > DNA <213> Artificial Sequence <400> 79 caatggctac aatgtctcca g 21 <210> 80 <211> 20 <212> DNA <213> Artificial Sequence <400> 80 ggtccctgat ggctacaatg 20 <210> 81 <211> 21 <212> DNA <213> Artificial Sequence <400> 81 tgatggttat agtgtctcca g 21 <210> 82 <211> 20 <212> DNA <213> Artificial Sequence <400> 82 ccgaatggct acaacgtctc 20 <210> 83 <211> 21 <212> DNA <213> Artificial Sequence <400> 83 gtctctcgaa aagagaagag g 21 <210> 84 <211> 20 <212> DNA <213> Artificial Sequence <400> 84 agtgtctctc gacaggcaca 20 <210> 85 <211> 21 <212> DNA < 213> Artificial Sequence <400> 85 gtgaaagagt ctaaacaggat 21 <210> 86 <211> 23 <212> DNA <213> Artificial Sequence <400> 86 cacagatagt aaatgacttt cag 23 <210> 87 <211> 21 <212> DNA < 213> Artificial Sequence <400> 87 gatgagtcag gaatgccaaa g 21 <210> 88 <211> 20 <212> DNA <213> A rtificial Sequence <400> 88 ctcaatgccc caagaacgca 20 <210> 89 <211> 20 <212> DNA <213> Artificial Sequence <400> 89 tctgaggtgc cccagaatct 20 <210> 90 <211> 20 <212> DNA <213> Artificial Sequence <400> 90 tctgcagaga ggctcaaagg 20 <210> 91 <211> 23 <212> DNA <213> Artificial Sequence <400> 91 ttcagtgact atcattctga act 23 <210> 92 <211> 21 <212> DNA <213> Artificial Sequence < 400> 92 attcttgggg cagaaagtcg a 21 <210> 93 <211> 19 <212> DNA <213> Artificial Sequence <400> 93 ccaggaggcc gaacacttc 19 <210> 94 <211> 21 <212> DNA <213> Artificial Sequence <400 > 94 aacaggtatg cccaaggaaa g 21 <210> 95 <211> 19 <212> DNA <213> Artificial Sequence <400> 95 cacccgaggt cgctgtgtt 19 <210> 96 <211> 21 <212> DNA <213> Artificial Sequence <400> 96 gctgtgtttg agccatcaga a 21 <210> 97 <211> 27 <212> PRT <213> Homo sapiens <400> 97 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Val Gly Asn Gly Tyr 1 5 10 15 Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 <210> 98 <211> 30 <212> PRT <213> Homo sapiens <400> 98 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Asp Glu Ala Ile 1 5 10 15 Thr Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 99 <211> 31 <212> PRT < 213> Homo sapiens <400> 99 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Asp Pro Thr Gly Arg 1 5 10 15 His Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 100 <211> 29 <212> PRT <213> Homo sapiens <400> 100 Glu Asp Ser Ser Phe Tyr Ile Cys Arg Pro Ser Ser Gly Ser Tyr Asn 1 5 10 15 Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 101 <211> 34 <212> PRT <213> Homo sapiens <400> 101 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Leu Ala Gly Thr 1 5 10 15 Ser Ser Ser Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr 20 25 30 Val Leu <210> 102 <211> 34 <212> PRT <213> Homo sapiens <400> 102 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ile Gly Thr Thr Ser Gly 1 5 10 15 Ser His Arg Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr 20 25 30 Val Leu <210> 103 <211> 26 <212> PRT <213> Homo sapie ns <400> 103 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ser Gly Glu Thr Gln Tyr 1 5 10 15 Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 104 <211> 30 <212> PRT < 213> Homo sapiens <400> 104 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Pro Ser Gly Arg Ala Gln 1 5 10 15 Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 105 < 211> 28 <212> PRT <213> Homo sapiens <400> 105 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Val Gly Gly Gln Ser Tyr Glu 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 106 <211> 28 <212> PRT <213> Homo sapiens <400> 106 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Ala Gln Asp Val Tyr Glu 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 107 <211> 29 <212> PRT <213> Homo sapiens <400> 107 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Asp Thr Arg Gly Met Asn Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 108 <211> 31 <212> PRT <213> Homo sapiens <400> 108 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Al a Asp Arg Tyr 1 5 10 15 Asn Thr Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 109 <211> 29 <212> PRT <213> Homo sapiens <400> 109 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Glu Thr Ser Asn Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 110 <211> 28 <212> PRT <213> Homo sapiens < 400> 110 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Phe Ser Ser Gly Gly 1 5 10 15 Tyr Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 <210> 111 <211> 30 <212> PRT < 213> Homo sapiens <400> 111 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Asn Leu Ala Gly Val Gly 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 112 < 211> 30 <212> PRT <213> Homo sapiens <400> 112 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Val Ser Gly Arg Thr 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 113 <211> 31 <212> PRT <213> Homo sapiens <400> 113 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Tyr Thr Ser Gly Arg Ser 1 5 10 15 Lys A sn Ile Gln Tyr Phe Gly Ala Gly Thr Arg Leu Ser Val Leu 20 25 30 <210> 114 <211> 29 <212> PRT <213> Homo sapiens <400> 114 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Leu Ala Val Glu 1 5 10 15 Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 115 <211> 31 <212> PRT <213> Homo sapiens <400> 115 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Pro Ile Val Gly Gly 1 5 10 15 Ser His Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 116 <211> 30 <212> PRT <213> Homo sapiens < 400> 116 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Arg Gly Pro Gly Thr Ser 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 117 <211> 30 <212 > PRT <213> Homo sapiens <400> 117 Glu Asp Ser Ser Phe Tyr Ile Cys Ser Ala Thr Ser Ser Gly Ser Ser 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210 > 118 <211> 29 <212> PRT <213> Homo sapiens <400> 118 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Glu Asp Val Gly Thr 1 5 10 15 Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 <210> 119 <211> 32 <212> PRT <213> Homo sapiens <400> 119 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Thr Ser Gly Gly 1 5 10 15 Ala Trp Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 120 <211> 29 <212> PRT <213> Homo sapiens <400> 120 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ile Lys Thr Asp Pro Arg Arg 1 5 10 15 Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 121 <211> 28 <212> PRT <213> Homo sapiens <400> 121 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Arg Ser Gly Gly Gly Glu 1 5 10 15 Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 <210> 122 <211> 33 <212> PRT <213> Homo sapiens <400 > 122 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Ala Gln Gly 1 5 10 15 Gln Gly Lys Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Leu <210> 123 <211> 31 <212> PRT <213> Homo sapiens <400> 123 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Arg Leu Ala Gly Ala Phe 1 5 10 15 Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 124 <211> 28 <212> PRT <213> Homo sapiens <400> 124 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Ile Thr Asp Thr 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 125 <211> 32 <212> PRT <213> Homo sapiens <400> 125 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Gly Gly Gly 1 5 10 15 Arg Val Ala Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 126 <211> 31 <212> PRT <213> Homo sapiens <400> 126 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Gly Gln Glu 1 5 10 15 Asn Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 127 <211> 32 <212> PRT <213> Homo sapiens < 400> 127 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Arg Ala Ser Gly 1 5 10 15 Arg Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 128 <211> 27 <212> PRT <213> Homo sapiens <400> 128 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Glu Leu Gly Phe 1 5 10 15 Thr Phe Gly Ser Gly Thr Arg Leu Thr Val V al 20 25 <210> 129 <211> 29 <212> PRT <213> Homo sapiens <400> 129 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Phe Gly Gly Glu 1 5 10 15 Lys Leu Phe Phe Gly Ser Gly Thr Gln Leu Ser Val Leu 20 25 <210> 130 <211> 29 <212> PRT <213> Homo sapiens <400> 130 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Arg Asp Gln 1 5 10 15 Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 <210> 131 <211> 30 <212> PRT <213> Homo sapiens <400> 131 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Pro Gln Ser Gly His Ala 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 132 <211> 28 <212> PRT <213> Homo sapiens <400> 132 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Phe Thr Asp Thr 1 5 10 15 Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 133 <211> 26 <212> PRT <213> Homo sapiens <400> 133 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Pro Pro Gln Tyr 1 5 10 15 Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 134 <211> 27 <212> PRT <213> Homo sapiens <4 00> 134 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Lys Trp Thr Gln 1 5 10 15 Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 <210> 135 <211> 27 <212> PRT <213 > Homo sapiens <400> 135 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Arg Glu Gly Gly Gln 1 5 10 15 Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 <210> 136 <211> 31 <212 > PRT <213> Homo sapiens <400> 136 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Gly Ser Leu Ser 1 5 10 15 Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 < 210> 137 <211> 30 <212> PRT <213> Homo sapiens <400> 137 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Ala Gln Gly Ala 1 5 10 15 Ala Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 138 <211> 30 <212> PRT <213> Homo sapiens <400> 138 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Gly Ser Leu Val Asp 1 5 10 15 Gln Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 30 <210> 139 <211> 30 <212> PRT <213> Homo sapiens <400> 139 Gly Asp Ser Ala Met Tyr Leu Cys A la Ser Ser Gln Gln Gly Gly Gly 1 5 10 15 Asn Thr Ile Tyr Phe Gly Glu Gly Ser Trp Leu Thr Val Val 20 25 30 <210> 140 <211> 31 <212> PRT <213> Homo sapiens <400> 140 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Ser Leu Val 1 5 10 15 Asp Gln Pro Gln His Phe Gly Asp Gly Thr Arg Leu Ser Ile Leu 20 25 30 <210> 141 <211> 30 <212> PRT <213> Homo sapiens <400> 141 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ile Gly Ala Ala 1 5 10 15 His Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 142 <211> 27 <212> PRT <213> Homo sapiens <400> 142 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Asp Glu Arg Glu 1 5 10 15 Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 <210> 143 <211> 31 <212> PRT <213> Homo sapiens <400> 143 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Ser Thr Arg 1 5 10 15 Gly Asp Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 144 <211> 31 <212> PRT <213> Homo sapiens <400> 144 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Gln Gly Gly Gly Gly 1 5 10 15 Thr Gly Glu Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 30 <210> 145 <211> 33 <212> PRT <213> Homo sapiens <400> 145 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Leu Val Arg 1 5 10 15 Asp Tyr Ser Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Thr <210> 146 <211> 30 <212> PRT <213> Homo sapiens <400> 146 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Thr Gly Gly Gly 1 5 10 15 Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 147 <211> 30 <212> PRT <213> Homo sapiens <400> 147 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Thr Thr Asp Ala 1 5 10 15 Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 148 <211> 31 <212> PRT <213> Homo sapiens <400> 148 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ala Ile Ala Gly Ala 1 5 10 15 Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 149 <211> 30 <212> PRT <213> Homo sapiens <400> 149 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ala Gl y Gln Gly Ser 1 5 10 15 Thr Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 150 <211> 31 <212> PRT <213> Homo sapiens <400> 150 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Leu Val Gly Gly 1 5 10 15 Arg Gly Glu Ala Phe Phe Gly Gln Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 151 <211> 30 <212> PRT <213> Homo sapiens <400> 151 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Leu Gly Gln Asn 1 5 10 15 Tyr Gly Tyr Thr Phe Gly Ser Gly Thr Arg Leu Thr Val Val 20 25 30 <210> 152 <211> 27 <212> PRT <213> Homo sapiens <400> 152 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Val Asp Thr Ile 1 5 10 15 Tyr Phe Gly Glu Gly Ser Trp Leu Thr Val Val 20 25 <210> 153 <211> 32 <212> PRT <213> Homo sapiens <400> 153 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Pro Gly Thr Ser Gly 1 5 10 15 Ser Tyr Asn Glu Gln Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu 20 25 30 <210> 154 <211> 31 <212> PRT <213> Homo sapiens <400> 154 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Phe Gly Arg Gly Asn 1 5 10 15 Thr Gly Glu Leu Phe Phe Gly Glu Gly Ser Arg Leu Thr Val Leu 20 25 30 <210> 155 <211> 33 <212> PRT <213> Homo sapiens <400> 155 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Ala Ile Arg Gly 1 5 10 15 Ala Ser Thr Asp Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val 20 25 30 Leu <210> 156 <211> 31 <212> PRT <213> Homo sapiens <400 > 156 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Thr Ser Gly 1 5 10 15 Ala Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 <210> 157 <211> 31 <212 > PRT <213> Homo sapiens <400> 157 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Val Thr Pro 1 5 10 15 Gln Glu Thr Gln Tyr Phe Gly Pro Gly Thr Arg Leu Leu Val Leu 20 25 30 < 210> 158 <211> 36 <212> PRT <213> Homo sapiens <400> 158 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ile Pro Arg Leu Gly 1 5 10 15 Gln Gly Ala Ser Gly Ala Asn Val Leu Thr Phe Gly Ala Gly Ser Arg 20 25 30 Leu Thr Val Leu 35 <210> 159 <211> 30 <212> PRT <213> Homo sapiens <400> 159 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Leu Gly Thr Gly Gly 1 5 10 15 Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30 <210> 160 <211> 32 <212> PRT <213> Homo sapiens <400> 160 Gly Asp Ser Ala Met Tyr Leu Cys Ala Ser Ser Ser Ile Pro Gly Leu 1 5 10 15 Arg Asn Tyr Glu Gln Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Thr 20 25 30
【図１】 川崎病患者（２２名の）発症期、γグロブリ
ン投与期、及び回復期の末梢血におけるＶα鎖サブファ
ミリーの使用頻度（％）を示すグラフ。FIG. 1 is a graph showing the frequency (%) of use of the Vα chain subfamily in peripheral blood of Kawasaki disease patients (22 patients) during onset, γ globulin administration, and recovery.
【図２】 川崎病患者（２２名の）発症期、γグロブリ
ン投与期、及び回復期の末梢血におけるＶβ鎖サブファ
ミリーの使用頻度（％）を示すグラフ。FIG. 2 is a graph showing the frequency of use (%) of the Vβ chain subfamily in peripheral blood of Kawasaki disease patients (22 patients) during onset, γ globulin administration, and recovery.
【図３】 川崎病患者２２名の治療前と回復期、発熱を
伴う川崎病以外の疾患群の子供１４名、及び健常な大人
１０名についてＴＣＲＶβ鎖のＶβ２.１及びＶβ６.５
の発現頻度を示すグラフ。FIG. 3: Vβ2.1 and Vβ6.5 of TCRVβ chain in 22 children with Kawasaki disease before and after treatment, 14 children in a group of diseases other than Kawasaki disease with fever, and 10 healthy adults.
Is a graph showing the expression frequency.
───────────────────────────────────────────────────── フロントページの続き Ｆターム(参考） 2G045 AA02 AA25 AA35 CA18 CA25 DA13 FB01 FB02 GA01 4B024 AA01 AA11 CA04 CA09 CA11 DA03 EA04 HA11 4B063 QA01 QA19 QQ03 QQ52 QR31 QR32 QR55 QR62 QR84 QS02 QS16 QS25 QX01 QX02 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G045 AA02 AA25 AA35 CA18 CA25 DA13 FB01 FB02 GA01 4B024 AA01 AA11 CA04 CA09 CA11 DA03 EA04 HA11 4B063 QA01 QA19 QQ03 QQ52 QR31 QR32 QR55 QR62 QR84 QS02 QS16 QQSQ
Claims (4)
β６.５及びＶβ２.１陽性Ｔ細胞数の増加を指標とする
川崎病因子の検出法。1. An increase in the number of Vβ6.5-positive T cells, or
A method for detecting Kawasaki disease factor using an increase in the number of β6.5 and Vβ2.1 positive T cells as an index.
ＲＶ領域の遺伝子群を増幅し、ＴＣＲＶサブファミリー
の発現頻度を検出し、ＴＣＲＶβ６.５及びＴＣＲＶβ
２.１サブファミリーの発現頻度の有意な増大を指標と
する請求項１記載の検出法。2. TC in peripheral blood of a patient suspected of Kawasaki disease
The genes in the RV region were amplified, the frequency of expression of the TCRV subfamily was detected, and TCRVβ6.5 and TCRVβ
2. The detection method according to claim 1, wherein a significant increase in the expression frequency of the 2.1 subfamily is used as an index.
ＲＶ領域遺伝子群をアダプター付加ＰＣＲ法で増幅し、
少なくとも配列番号１０及び１１に記載の塩基配列で示
されるオリゴヌクレオチドプローブを含むＴＣＲＶサブ
ファミリーに特異的なオリゴヌクレオチドプローブを用
いるリバース・ドット・ブロット法により、ＴＣＲＶβ
サブファミリーの発現頻度を検出し、ＴＣＲＶβ６.５
サブファミリー、又はＴＣＲＶβ６.５及びＴＣＲＶβ
２.１サブファミリーの発現頻度が、他のサブファミリ
ーの発現頻度よりも有意に高いことを指標とする請求項
１又は２記載の方法。3. TC in peripheral blood of a patient suspected of Kawasaki disease
The RV region genes are amplified by an adapter-added PCR method,
By reverse dot blotting using an oligonucleotide probe specific to the TCRV subfamily containing at least the oligonucleotide probes represented by the nucleotide sequences of SEQ ID NOS: 10 and 11, TCRVβ
The expression frequency of the subfamily was detected, and TCRVβ6.5 was detected.
Subfamily, or TCRVβ6.5 and TCRVβ
3. The method according to claim 1 or 2, wherein the expression frequency of the 2.1 subfamily is significantly higher than the expression frequency of the other subfamilies.
β６.５陽性ｃＤＮＡクローンのＣＤＲ３領域の塩基配
列を決定し、ＴＣＲＶβ２.１及びＴＣＲＶβ６.５陽性
Ｔ細胞数の増加がポリクローナルであることを確認する
ことをも含む請求項１〜３のいずれかに記載の方法。4. The method of claim 1, further comprising the steps of:
4. The method according to claim 1, further comprising determining the nucleotide sequence of the CDR3 region of the β6.5-positive cDNA clone, and confirming that the increase in the number of TCRVβ2.1 and TCRVβ6.5-positive T cells is polyclonal. The described method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10341661A JP2000157297A (en) | 1998-12-01 | 1998-12-01 | Method for detecting factor of kawasaki disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10341661A JP2000157297A (en) | 1998-12-01 | 1998-12-01 | Method for detecting factor of kawasaki disease |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2000157297A true JP2000157297A (en) | 2000-06-13 |
Family
ID=18347821
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10341661A Pending JP2000157297A (en) | 1998-12-01 | 1998-12-01 | Method for detecting factor of kawasaki disease |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2000157297A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1517995A2 (en) * | 2002-07-03 | 2005-03-30 | Institute For Scientific Research, Inc. | Compositions and methods for the detection of human t cell receptor variable family gene expression |
| JP2007205842A (en) * | 2006-02-01 | 2007-08-16 | Kobeshi Chiiki Iryo Shinko Zaidan | Kawasaki disease determination method and kit for the same |
| WO2009019657A3 (en) * | 2007-08-06 | 2009-08-06 | Univ Cattolica Sacro Cuore | Means for diagnosis and treatment of rheumatoid arthritis |
| EP2281065A4 (en) * | 2008-04-16 | 2011-08-31 | Hudsonalpha Inst For Biotechnology | Method for evaluating and comparing immunorepertoires |
-
1998
- 1998-12-01 JP JP10341661A patent/JP2000157297A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1517995A2 (en) * | 2002-07-03 | 2005-03-30 | Institute For Scientific Research, Inc. | Compositions and methods for the detection of human t cell receptor variable family gene expression |
| EP1517995A4 (en) * | 2002-07-03 | 2007-07-11 | Inst Scient Res Inc | Compositions and methods for the detection of human t cell receptor variable family gene expression |
| JP2007205842A (en) * | 2006-02-01 | 2007-08-16 | Kobeshi Chiiki Iryo Shinko Zaidan | Kawasaki disease determination method and kit for the same |
| WO2009019657A3 (en) * | 2007-08-06 | 2009-08-06 | Univ Cattolica Sacro Cuore | Means for diagnosis and treatment of rheumatoid arthritis |
| EP2281065A4 (en) * | 2008-04-16 | 2011-08-31 | Hudsonalpha Inst For Biotechnology | Method for evaluating and comparing immunorepertoires |
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