JP2003287538A - Dna chip and dna testing method - Google Patents
Dna chip and dna testing methodInfo
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- JP2003287538A JP2003287538A JP2002285845A JP2002285845A JP2003287538A JP 2003287538 A JP2003287538 A JP 2003287538A JP 2002285845 A JP2002285845 A JP 2002285845A JP 2002285845 A JP2002285845 A JP 2002285845A JP 2003287538 A JP2003287538 A JP 2003287538A
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- dna
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、特定の塩基配列を
有するＤＮＡを検出するＤＮＡチップを用いたＤＮＡ検
査方法に関するものである。TECHNICAL FIELD The present invention relates to a DNA inspection method using a DNA chip for detecting DNA having a specific base sequence.
【０００２】[0002]
【従来の技術】近年、人の遺伝子構造がほぼ解明され、
医療などの応用のために解明された遺伝子の持つ機能の
調査，研究が本格化してきている。この遺伝子の機能解
明のために、ＤＮＡチップが用いられている。ＤＮＡ
は、Ａ（アデニン），Ｔ（チミン），Ｃ（シトシン），
Ｇ（グアニン）の４つの塩基により形成された２本の分
子鎖が、螺旋状に結合して形成されたものである。４つ
の塩基のうち、結合可能な組み合わせは、ＡとＴ、Ｃと
Ｇである。2. Description of the Related Art In recent years, the genetic structure of humans has been elucidated,
Research and research into the functions of genes that have been elucidated for medical and other applications are in full swing. A DNA chip is used to elucidate the function of this gene. DNA
Is A (adenine), T (thymine), C (cytosine),
Two molecular chains formed by the four bases of G (guanine) are formed by helically binding. Among the four bases, combinations capable of binding are A and T and C and G.
【０００３】ＤＮＡチップは、ガラスやシリコンなどの
基板上に、高密度にＤＮＡ分子の断片を固定したもので
ある。例えば、検体となるＤＮＡを被検者の血液から抽
出し、抽出した溶液中のＤＮＡを１本鎖に分解した後、
抽出した溶液をＤＮＡチップの表面に滴下し、ＤＮＡチ
ップ上のＤＮＡ断片との結合を調べて検体の種類を判定
する。A DNA chip is one in which fragments of DNA molecules are immobilized at high density on a substrate such as glass or silicon. For example, a sample DNA is extracted from the blood of a subject, and the DNA in the extracted solution is decomposed into single strands,
The extracted solution is dropped on the surface of the DNA chip, and the binding to the DNA fragment on the DNA chip is examined to determine the type of sample.
【０００４】現在、開発され一部実用化されているＤＮ
Ａチップを用いた検出方式として、レーザを照射して蛍
光を測定する蛍光検出方式がある。この方式では、検体
となるＤＮＡに予め蛍光色素で標識をつけ、ＤＮＡチッ
プ上のＤＮＡ断片に結合した検体ＤＮＡの有無を、レー
ザ光照射による蛍光色素の発光により検出している（非
特許文献１参照）。Currently developed and partially put into practical use
As a detection method using the A chip, there is a fluorescence detection method in which a laser is irradiated to measure fluorescence. In this method, a sample DNA is labeled with a fluorescent dye in advance, and the presence or absence of the sample DNA bound to the DNA fragment on the DNA chip is detected by the emission of the fluorescent dye by laser light irradiation (Non-Patent Document 1). reference).
【０００５】[0005]
【非特許文献１】原田 学，佐藤 高遠，米田 英克、
「ＤＮＡチップの現状と展望」、応用物理、第６９巻、
第１２号（２０００）[Non-Patent Document 1] Manabu Harada, Takatoshi Sato, Hidekatsu Yoneda,
"Current status and prospects of DNA chips", Applied Physics, Volume 69,
No. 12 (2000)
【０００６】[0006]
【発明が解決しようとする課題】しかしながら、前述し
たような、従来の技術では、レーザ照射装置など大がか
りな装置が必要となり、システムが高価なものとなる。
また、定量的に検出することが容易ではなかった。本発
明は、以上のような問題点を解消するためになされたも
のであり、容易にＤＮＡの検査ができるようにすること
を目的とする。However, in the conventional technique as described above, a large-scale device such as a laser irradiation device is required, and the system becomes expensive.
Moreover, it was not easy to detect quantitatively. The present invention has been made to solve the above problems, and an object of the present invention is to enable easy DNA inspection.
【０００７】[0007]
【課題を解決するための手段】本発明のＤＮＡチップ
は、所定の間隔で各々分離した複数の例えば凸部や凹部
からなる島部を備えた水晶基板と、生化学物質のＤＮＡ
に特有な塩基配列から構成されて各々の島部の上に固定
された複数のＤＮＡ断片とから構成されたものである。
このＤＮＡチップでは、各島部における共振周波数の変
化により、各島部上に固定されているＤＮＡ断片に結合
したＤＮＡの有無を検出する。The DNA chip of the present invention comprises a quartz substrate having a plurality of islands, each of which is composed of, for example, a convex portion and a concave portion, which are separated at a predetermined interval, and a DNA of a biochemical substance.
It is composed of a plurality of DNA fragments which are composed of a unique nucleotide sequence and are fixed on each island.
In this DNA chip, the presence or absence of DNA bound to the DNA fragment fixed on each island is detected by the change in resonance frequency at each island.
【０００８】上記ＤＮＡチップにおいて、島部上に金薄
膜を形成し、ＤＮＡ断片は、一端をＳＨ基に置換し、こ
のＳＨ基によりＤＮＡ断片を金薄膜に固定すればよい。
また、島部は、水晶基板の表面を親水性にする親水処理
を施した領域であってもよい。また、上記ＤＮＡチップ
において、複数の島部上に、各々異なる塩基配列のＤＮ
Ａ断片が固定されているようにしてもよい。In the above DNA chip, a gold thin film may be formed on the island portion, one end of the DNA fragment may be replaced with an SH group, and the DNA fragment may be fixed to the gold thin film by this SH group.
Further, the island portion may be a region that has been subjected to a hydrophilic treatment to make the surface of the quartz substrate hydrophilic. In addition, in the above-mentioned DNA chip, DNs having different base sequences are provided on a plurality of islands.
The A fragment may be fixed.
【０００９】また、本発明のＤＮＡ検査方法は、所定の
間隔で各々分離した複数の凸部や凹部などからなる島部
を備えた水晶基板を用意し、各々の島部上に生化学物質
のＤＮＡに特有な塩基配列から構成された所望のＤＮＡ
断片を固定し、ＤＮＡ断片が固定された各々島部の共振
周波数を測定して島部各々の第１の測定周波数とし、島
部上に固定されたＤＮＡ断片を、検体となるＤＮＡを含
んだ溶液中に所定時間接触させた後で乾燥し、各々の島
部の共振周波数を測定して島部各々の第２の測定周波数
とし、第２の測定周波数と第１の測定周波数との差によ
り、検体となるＤＮＡの中よりＤＮＡ断片と同じ塩基配
列を検出するようにしたものである。このＤＮＡ検査方
法によれば、各島部における第１の測定周波数と第２の
測定周波数との差により、各島部上に固定されているＤ
ＮＡ断片に結合したＤＮＡの有無を検出する。Further, in the DNA testing method of the present invention, a quartz substrate having an island portion composed of a plurality of convex portions and concave portions separated at a predetermined interval is prepared, and a biochemical substance is placed on each island portion. Desired DNA composed of a nucleotide sequence unique to DNA
The fragment was fixed, and the resonance frequency of each island where the DNA fragment was fixed was measured as the first measurement frequency of each island, and the DNA fragment fixed on the island contained the sample DNA. After being in contact with the solution for a predetermined time, it is dried, and the resonance frequency of each island is measured to obtain the second measurement frequency of each island, which is determined by the difference between the second measurement frequency and the first measurement frequency. The same base sequence as the DNA fragment is detected from the sample DNA. According to this DNA inspection method, D fixed on each island part is determined by the difference between the first measurement frequency and the second measurement frequency on each island part.
The presence or absence of DNA bound to the NA fragment is detected.
【００１０】[0010]
【発明の実施の形態】以下、本発明の実施の形態につい
て図を参照して説明する。図１は、本発明の実施の形態
におけるＤＮＡチップの構成を示す断面図（ａ）と平面
図（ｂ）である。このＤＮＡチップは、ＡＴカットの水
晶基板１０１上に、直径１ｍｍ程度，高さ１０μｍ程度
の複数の凸部（島部）１０２が、２ｍｍ間隔でマトリク
ス状に形成され、これら複数の凸部１０２上に形成され
た金薄膜１０３の表面に、所望とする複数のＤＮＡ断片
１０４が各々固定されているものである。BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is a sectional view (a) and a plan view (b) showing a configuration of a DNA chip according to an embodiment of the present invention. In this DNA chip, a plurality of convex portions (islands) 102 having a diameter of about 1 mm and a height of about 10 μm are formed in a matrix form at intervals of 2 mm on an AT-cut quartz crystal substrate 101. A plurality of desired DNA fragments 104 are immobilized on the surface of the gold thin film 103 formed in the above.
【００１１】凸部１０２の上へのＤＮＡ断片１０４の固
定は、つぎに示すようにする。まず、所望とするＤＮＡ
断片の一端がＳＨ基で置換された状態とする。次いで、
ＳＨ基で一端が置換されたＤＮＡ断片が分散している溶
媒中に、金薄膜１０３が各凸部１０２の表面に形成され
た水晶基板１０１を浸漬する。このことにより、金薄膜
１０３の上にＳＨ基が引き寄せられて固着する。この結
果、金薄膜１０３の表面にＳＨ基を介してＤＮＡ断片１
０４が固定された状態となる。この後、水晶基板１０１
は、純水で洗浄してから乾燥すればよい。The DNA fragment 104 is fixed on the convex portion 102 in the following manner. First, the desired DNA
It is assumed that one end of the fragment is replaced with the SH group. Then
The quartz substrate 101 on which the gold thin film 103 is formed on the surface of each convex portion 102 is immersed in a solvent in which a DNA fragment whose one end is replaced with an SH group is dispersed. As a result, the SH group is attracted and fixed on the gold thin film 103. As a result, the DNA fragment 1 is formed on the surface of the gold thin film 103 via the SH group.
04 is fixed. After this, the crystal substrate 101
May be washed with pure water and then dried.
【００１２】以下に、この実施の形態におけるＤＮＡ検
査方法について説明する。まず、ＤＮＡ断片１０４が固
定されていない状態の水晶基板１０１の各凸部１０２の
共振周波数（Ｆ0：基本振動数）を測定する。つぎに、
前述した方法により、所望のＤＮＡ断片１０４を各々異
なる凸部１０２上に固定し、水晶基板１０１を純水で洗
浄して乾燥した後、複数のＤＮＡ断片１０４が固定され
ている領域（島部）である各凸部１０２の共振周波数
（Ｆ1：第１の測定周波数）を測定する。The DNA inspection method according to this embodiment will be described below. First, the resonance frequency (F0: fundamental frequency) of each convex portion 102 of the quartz substrate 101 in which the DNA fragment 104 is not fixed is measured. Next,
According to the method described above, desired DNA fragments 104 are fixed on different convex portions 102 respectively, and the quartz substrate 101 is washed with pure water and dried, and then a plurality of DNA fragments 104 are fixed (islands). The resonance frequency (F1: first measurement frequency) of each convex portion 102 is measured.
【００１３】ここで、各凸部１０２上においてＤＮＡ断
片１０４が形成される領域の面積Ａ、凸部１０２部分の
厚さｔ、水晶の密度ρ、共振周波数の変化ΔＦ、上記面
積Ａの領域の上における質量変化Δｍ、とすると、これ
らの関係は、「−ΔＦ＝（Ｆ0・Δｍ）／（ρ・Ａ・
ｔ）」で示される。したがって、各凸部１０２に固定さ
れたＤＮＡ断片１０４によって、質量変化Δｍ1が生
じ、共振周波数の変化（ΔＦ01＝Ｆ0−Ｆ1）として検出
することができる。Here, the area A of the region where the DNA fragment 104 is formed on each convex portion 102, the thickness t of the convex portion 102 portion, the density ρ of the crystal, the change ΔF of the resonance frequency, and the area of the above area A Assuming that the mass change Δm above, the relationship between them is “−ΔF = (F0 · Δm) / (ρ · A ·
t) ”. Therefore, the DNA fragment 104 fixed to each convex portion 102 causes a mass change Δm1, which can be detected as a change in resonance frequency (ΔF01 = F0−F1).
【００１４】つぎに、ＤＮＡの２重螺旋構造が解ける程
度の温度とした状態で、検体となるＤＮＡを含んだ溶液
中にＤＮＡ断片１０４が固定された水晶基板１０１を所
定時間浸漬するなどにより、ＤＮＡ断片１０４部分を上
記溶液に接触させる。この後、溶液温度を低下させてＤ
ＮＡが螺旋構造をとる状態としてから水晶基板を溶液中
より引き上げ、これを純水で洗浄して乾燥し、各凸部１
０２の共振周波数（Ｆ2：第２の測定周波数）を測定す
る。Next, by immersing the quartz substrate 101, on which the DNA fragment 104 is fixed, in a solution containing the sample DNA, for a predetermined time, at a temperature at which the double helix structure of the DNA can be unraveled. A portion of the DNA fragment 104 is brought into contact with the above solution. After this, the solution temperature is lowered and D
After the NA has a spiral structure, the quartz substrate is pulled out of the solution, washed with pure water and dried to form each convex portion 1
The resonance frequency of 02 (F2: second measurement frequency) is measured.
【００１５】このとき、検体となるＤＮＡの塩基配列の
中に、ＤＮＡ断片１０４と同じ塩基配列が存在すると、
前述した水晶基板１０１の溶液に対する浸漬により、同
じ塩基配列が存在するＤＮＡはＤＮＡ断片１０４に結合
する。ＤＮＡ断片１０４に同じ塩基配列が存在するＤＮ
Ａが結合した場合、このＤＮＡ断片１０４が固定されて
いる凸部１０２上では、質量が増加（変化）したことに
なる。すなわち、検体となるＤＮＡと同じ塩基配列を有
するＤＮＡ断片１０４が固定された凸部１０２における
質量変化（Δｍ2）は、固定されたＤＮＡ断片１０４お
よびこのＤＮＡ断片１０４に結合したＤＮＡによるもの
である。At this time, if the same base sequence as the DNA fragment 104 exists in the base sequence of the sample DNA,
By immersing the quartz substrate 101 in the solution described above, DNAs having the same base sequence are bound to the DNA fragments 104. DN in which the same nucleotide sequence exists in the DNA fragment 104
When A is bound, the mass increases (changes) on the convex portion 102 to which the DNA fragment 104 is fixed. That is, the mass change (Δm 2) in the convex portion 102 to which the DNA fragment 104 having the same base sequence as the sample DNA is fixed is due to the fixed DNA fragment 104 and the DNA bound to this DNA fragment 104.
【００１６】したがって、水晶基板１０１を上記ＤＮＡ
を含んだ溶液に浸漬する前後で、Ｆ1とＦ2との差があれ
ば、質量変化（Δｍ1，Δｍ2）に差があることになる。
よって、Ｆ1とＦ2とに差が発生した凸部１０２上におい
て、先に固定してあったＤＮＡ断片１０４に検体中のＤ
ＮＡが結合することによる質量の変化（増加）が発生し
たことが検出できる。すなわち、ＤＮＡ断片同士の結合
による質量増加が、凸部１０２毎の共振周波数の測定に
よりＦ1とＦ2との差として検出されるので、どの凸部１
０２にＦ1とＦ2との差が生じたかを調べることにより、
検体となるＤＮＡの中よりＤＮＡ断片１０４と同じ塩基
配列を検出することが可能となる。Therefore, the crystal substrate 101 is replaced with the DNA described above.
If there is a difference between F1 and F2 before and after the immersion in the solution containing, there is a difference in the mass change (Δm1, Δm2).
Therefore, on the convex portion 102 where the difference between F1 and F2 occurs, the D fragment in the sample is fixed to the DNA fragment 104 previously fixed.
It can be detected that a change in mass (increase) due to NA binding occurs. That is, since the increase in mass due to the binding of DNA fragments to each other is detected as the difference between F1 and F2 by measuring the resonance frequency of each convex portion 102, which convex portion 1
By checking whether the difference between F1 and F2 occurred in 02,
It is possible to detect the same base sequence as the DNA fragment 104 from the sample DNA.
【００１７】このように、基本振動数Ｆ0をもとに、Ｆ1
とＦ2との差から、水晶基板１０１を上記ＤＮＡを含ん
だ溶液に浸漬する前後における各々の凸部１０２上の質
量変化を検出し、各々の凸部１０２におけるＦ1とＦ2と
の差（質量変化）の有無により検体となるＤＮＡの中よ
りＤＮＡ断片１０４と同じ塩基配列を検出する。また、
凸部１０２上のＤＮＡ断片１０４に結合した検体のＤＮ
Ａの質量は、Δｍ＝−（Ｆ2−Ｆ1）・（ρ・Ａ・ｔ）／
Ｆ0により求めることができる。また、Ｆ0とＦ1の差か
ら、各々の凸部１０２上に固定されているＤＮＡ断片１
０４の質量も得られるので、１個のＤＮＡ断片の質量が
判明していれば、凸部１０２上に固定されているＤＮＡ
断片１０４の数も求めることができる。Thus, based on the fundamental frequency F0, F1
From the difference between F1 and F2, the change in mass on each convex portion 102 before and after the crystal substrate 101 is immersed in the solution containing the above DNA is detected, and the difference between F1 and F2 at each convex portion 102 (change in mass) ), The same base sequence as the DNA fragment 104 is detected from the sample DNA. Also,
DN of sample bound to DNA fragment 104 on convex portion 102
The mass of A is Δm =-(F2-F1) · (ρ · A · t) /
It can be obtained by F0. Further, from the difference between F0 and F1, the DNA fragment 1 fixed on each convex portion 102
Since the mass of 04 is also obtained, if the mass of one DNA fragment is known, the DNA fixed on the convex portion 102 can be obtained.
The number of fragments 104 can also be determined.
【００１８】ここで、凸部１０２の共振周波数の測定に
ついて簡単に説明する。図２に示すように、下部電極２
０１上に水晶基板１０１を載置し、水晶基板１０１上に
上部電極２０２が複数設けられた上部電極固定絶縁基板
２０３を対向配置し、上部電極２０２の各々が、各凸部
１０２上に配置された状態とする。なお、下部電極２０
１は、水晶基板１０１の裏面全域にわたる大きさに形成
され、上部電極２０２は、凸部１０２と同一の間隔で、
上部電極固定絶縁基板２０３にマトリクス状に配置され
ている。Here, the measurement of the resonance frequency of the convex portion 102 will be briefly described. As shown in FIG. 2, the lower electrode 2
01, the crystal substrate 101 is placed on the crystal substrate 101, and the upper electrode fixing insulating substrate 203 having a plurality of upper electrodes 202 provided on the crystal substrate 101 is opposed to each other. Each of the upper electrodes 202 is disposed on each convex portion 102. It will be in a state of The lower electrode 20
1 is formed to have a size over the entire back surface of the quartz substrate 101, and the upper electrodes 202 are arranged at the same intervals as the convex portions 102.
The upper electrode fixed insulating substrate 203 is arranged in a matrix.
【００１９】以上に説明したように下部電極２０１およ
び上部電極２０２を配置したら、測定対象の凸部１０２
上に配置されている上部電極２０２と下部電極２０１と
の間に、発振回路２０４より所定の周波数の信号を供給
し、周波数カウンタ２０５により共振周波数を計測す
る。When the lower electrode 201 and the upper electrode 202 are arranged as described above, the convex portion 102 to be measured is placed.
A signal having a predetermined frequency is supplied from the oscillation circuit 204 between the upper electrode 202 and the lower electrode 201 arranged above, and the resonance frequency is measured by the frequency counter 205.
【００２０】また、図３に示すように、各上部電極２０
２より各々配線を取り出し、切り替え器３０１により適
宜切り替えながら、各凸部１０２の共振周波数を逐次測
定するようにしてもよい。また、各々の凸部１０２上
に、各々異なる塩基配列のＤＮＡ断片を固定し、複数種
類の検体が同時に測定できるようにしてもよい。Further, as shown in FIG. 3, each upper electrode 20
It is also possible to take out each wiring from No. 2 and sequentially measure the resonance frequency of each convex portion 102 while appropriately switching by the switch 301. Further, DNA fragments having different base sequences may be fixed on each convex portion 102 so that a plurality of types of samples can be simultaneously measured.
【００２１】なお、上述した実施の形態では、水晶基板
上に、所定の間隔で各々分離した複数の凸部を備えるよ
うにしたが、これに限るものではない。例えば、図４に
示すように、水晶基板４０１の上に所定の間隔で各々分
離した複数の凹部４０２を備え、各々の凹部４０２の底
面上に金薄膜４０３を形成し、金薄膜４０３の表面にＤ
ＮＡ断片１０４を固定するようにしてもよい。In the above-described embodiment, the quartz substrate is provided with a plurality of convex portions separated by a predetermined distance, but the present invention is not limited to this. For example, as shown in FIG. 4, a plurality of recesses 402 are provided on a quartz substrate 401, each recess 402 being separated by a predetermined distance, and a gold thin film 403 is formed on the bottom surface of each recess 402. D
The NA fragment 104 may be fixed.
【００２２】ところで、上述した実施の形態では、水晶
基板に金属膜を形成してこの上にＤＮＡ断片を固定する
ようにしたが、図５に示すように、水晶基板に直接ＤＮ
Ａ断片を固定してもよい。図５に示すＤＮＡチップは、
ＡＴカットの水晶基板１０１上に、直径１ｍｍ程度の円
形領域の親水処理部５０２が２ｍｍ間隔でマトリクス状
に形成され、これら複数の親水処理部５０２の上に、所
望とする複数のＤＮＡ断片１０４が各々固定されている
ものである。By the way, in the above-mentioned embodiment, the metal film is formed on the crystal substrate and the DNA fragments are fixed on the metal film. However, as shown in FIG.
The A fragment may be fixed. The DNA chip shown in FIG.
On the AT-cut quartz crystal substrate 101, hydrophilic regions 502 having a circular area with a diameter of about 1 mm are formed in a matrix at intervals of 2 mm, and a plurality of desired DNA fragments 104 are formed on the plurality of hydrophilic regions 502. Each is fixed.
【００２３】親水処理部５０２上へのＤＮＡ断片１０４
の固定は、親水処理がされた部分とＤＮＡは容易に結合
することから、つぎに示すようにする。所望とするＤＮ
Ａ断片が分散している溶媒中に、親水処理部５０２が形
成された水晶基板１０１を浸漬する。このことにより、
親水処理部５０２の上にＤＮＡ断片１０４が引き寄せら
れて固着する。この結果、親水処理部５０２の表面にＤ
ＮＡ断片１０４が固定された状態となる。この後、水晶
基板１０１は、純水で洗浄してから乾燥すればよい。DNA fragment 104 on hydrophilic treatment section 502
The immobilization is carried out as follows, because the hydrophilically treated portion and the DNA are easily bound to each other. Desired DN
The quartz substrate 101 on which the hydrophilic treatment portion 502 is formed is immersed in the solvent in which the A fragment is dispersed. By this,
The DNA fragment 104 is attracted and fixed on the hydrophilic treatment portion 502. As a result, D is formed on the surface of the hydrophilic treatment portion 502.
The NA fragment 104 is fixed. After that, the quartz substrate 101 may be washed with pure water and then dried.
【００２４】図５のＤＮＡチップにおいても、つぎに示
すことにより、親水処理部５０２における水晶基板１０
１の共振周波数を測定することができる。まず、下部電
極２０１上に水晶基板１０１を載置し、水晶基板１０１
上に上部電極２０２が複数設けられた上部電極固定絶縁
基板２０３を対向配置し、上部電極２０２の各々が、所
定の間隔で各親水処理部５０２上に配置された状態とす
る。なお、下部電極２０１は、水晶基板１０１の裏面全
域にわたる大きさに形成され、上部電極２０２は、親水
処理部５０２と同一の間隔で、上部電極固定絶縁基板２
０３にマトリクス状に配置されている。Also in the DNA chip of FIG. 5, the quartz substrate 10 in the hydrophilic treatment portion 502 is shown as follows.
A resonant frequency of 1 can be measured. First, the quartz substrate 101 is placed on the lower electrode 201, and the quartz substrate 101
The upper electrode fixed insulating substrate 203 on which a plurality of upper electrodes 202 are provided is arranged so as to face each other, and each of the upper electrodes 202 is arranged on each hydrophilic treatment portion 502 at a predetermined interval. The lower electrode 201 is formed to have a size over the entire back surface of the quartz substrate 101, and the upper electrodes 202 are arranged at the same intervals as the hydrophilic treatment portion 502, and the upper electrode fixed insulating substrate 2 is provided.
03 are arranged in a matrix.
【００２５】以上に説明したように下部電極２０１およ
び上部電極２０２を配置したら、測定対象の親水処理部
５０２上に配置されている上部電極２０２と下部電極２
０１との間に、発振回路２０４より所定の周波数の信号
を供給し、周波数カウンタ２０５により共振周波数を計
測する。このようにして、親水処理部５０２の領域の共
振周波数を測定し、また、親水処理部５０２におけるＤ
ＮＡ断片１０４の有無およびＤＮＡ断片１０４に同じ塩
基配列が存在する検体のＤＮＡが結合したの各々におい
て共振周波数を測定すれば、前述と同様にＤＮＡの測定
検査が行える。After the lower electrode 201 and the upper electrode 202 are arranged as described above, the upper electrode 202 and the lower electrode 2 arranged on the hydrophilic treatment section 502 to be measured.
01, a signal of a predetermined frequency is supplied from the oscillation circuit 204, and the resonance frequency is measured by the frequency counter 205. In this way, the resonance frequency in the area of the hydrophilic processing section 502 is measured, and D in the hydrophilic processing section 502 is measured.
If the resonance frequency is measured for each of the presence or absence of the NA fragment 104 and the DNA of the sample in which the same base sequence exists in the DNA fragment 104, the measurement test of the DNA can be performed as described above.
【００２６】ところで、図５に示す実施の形態において
も、図１に示したように、水晶基板１０１に複数の凸部
を設け、この凸部の上面に親水処理を施し、親水処理さ
れた凸部の上面にＤＮＡ断片を固定するようにしてもよ
い。また、隣り合う親水処理部５０２の間に溝を設け、
親水処理部５０２と他の部分とに段差を設けるようにし
てもよい。これらのように、段差を設けることで、ＤＮ
Ａ断片が固定されている島部が、段差が無く平坦な状態
より発振しやすい状態となる。By the way, in the embodiment shown in FIG. 5 as well, as shown in FIG. 1, a plurality of convex portions are provided on the quartz substrate 101, and the upper surfaces of the convex portions are subjected to hydrophilic treatment, and the convex portions subjected to hydrophilic treatment are provided. You may make it fix a DNA fragment on the upper surface of a part. Further, a groove is provided between the adjacent hydrophilic treatment parts 502,
You may make it provide the level difference between the hydrophilic treatment part 502 and another part. By providing a step like this, DN
The island portion to which the A fragment is fixed becomes easier to oscillate than the flat state where there is no step.
【００２７】[0027]
【発明の効果】以上説明したように、本発明によれば、
水晶基板上に複数の島部を形成し、各島部における第１
の測定周波数と第２の測定周波数との差により、各島部
上に固定されているＤＮＡ断片に結合したＤＮＡの有無
を検出するようにしたので、ＤＮＡの検査がより容易に
行えるようになるというすぐれた効果が得られる。As described above, according to the present invention,
A plurality of islands are formed on a quartz substrate, and the first of each island is formed.
Since the presence or absence of DNA bound to the DNA fragment fixed on each island is detected by the difference between the measurement frequency and the second measurement frequency, it becomes easier to inspect the DNA. That is an excellent effect.
【図１】 本発明の実施の形態におけるＤＮＡチップの
構成例を示す概略的な断面図（ａ）と平面図（ｂ）であ
る。FIG. 1 is a schematic cross-sectional view (a) and a plan view (b) showing a configuration example of a DNA chip according to an embodiment of the present invention.
【図２】 本発明の実施の形態におけるＤＮＡ検査方法
を説明するための概略的な構成図である。FIG. 2 is a schematic configuration diagram for explaining a DNA inspection method according to an embodiment of the present invention.
【図３】 本発明の他の形態におけるＤＮＡ検査方法を
説明するための概略的な構成図である。FIG. 3 is a schematic configuration diagram for explaining a DNA inspection method according to another embodiment of the present invention.
【図４】 本発明の他の形態におけるＤＮＡチップの構
成例を示す概略的な断面図である。FIG. 4 is a schematic cross-sectional view showing a configuration example of a DNA chip according to another embodiment of the present invention.
【図５】 本発明の他の実施の形態におけるＤＮＡチッ
プの構成例を示す概略的な断面図である。FIG. 5 is a schematic cross-sectional view showing a configuration example of a DNA chip according to another embodiment of the present invention.
【符号の説明】
１０１…水晶基板、１０２…凸部（島部）、１０３…金
薄膜、１０４…ＤＮＡ断片。[Explanation of Codes] 101 ... Quartz substrate, 102 ... Convex part (island part), 103 ... Gold thin film, 104 ... DNA fragment.
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） Ｇ０１Ｎ 37/00 １０２ Ｃ１２Ｎ 15/00 Ｆ Ｆターム(参考） 4B024 AA11 CA01 CA09 HA14 4B029 AA07 BB20 CC03 FA15 4B063 QA01 QA13 QA18 QQ42 QR32 QR55 QR84 QS15 QS32 QS39 QX04 ─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. 7 Identification code FI theme code (reference) G01N 37/00 102 C12N 15/00 FF term (reference) 4B024 AA11 CA01 CA09 HA14 4B029 AA07 BB20 CC03 FA15 4B063 QA01 QA13 QA18 QQ42 QR32 QR55 QR84 QS15 QS32 QS39 QX04
Claims (9)
備えた水晶基板と、 生化学物質のＤＮＡに特有な塩基配列から構成されて各
々の前記島部の上に固定された複数のＤＮＡ断片とから
構成されたことを特徴とするＤＮＡチップ。1. A quartz substrate having a plurality of islands each separated at a predetermined interval, and a plurality of bases unique to DNA of a biochemical substance and fixed on the islands. A DNA chip comprising a DNA fragment.
れていることを特徴とするＤＮＡチップ。2. The DNA chip according to claim 1, wherein a gold thin film is formed on the island portion, one end of the DNA fragment is replaced with an SH group, and the DNA fragment is formed by the SH group. A DNA chip characterized by being fixed to.
理を施した領域であることを特徴とするＤＮＡチップ。3. The DNA chip according to claim 1, wherein the island portion is a region subjected to a hydrophilic treatment to make the surface of the quartz substrate hydrophilic.
ＮＡチップにおいて、 複数の前記島部上に、各々異なる塩基配列のＤＮＡ断片
が固定されていることを特徴とするＤＮＡチップ。4. D according to any one of claims 1 to 3.
In the NA chip, a DNA chip in which DNA fragments having different base sequences are fixed on the plurality of islands.
ＮＡチップにおいて、 前記島部は、前記水晶基板上に形成された凸部であるこ
とを特徴とするＤＮＡチップ。5. D according to any one of claims 1 to 4.
In the NA chip, the island part is a convex part formed on the crystal substrate, and the DNA chip is characterized.
ＮＡチップにおいて、 前記島部は、前記水晶基板上に形成された凹部であるこ
とを特徴とするＤＮＡチップ。6. D according to any one of claims 1 to 4.
In the NA chip, the island part is a recess formed on the crystal substrate, and the DNA chip is characterized.
備えた水晶基板を用意し、 各々の前記島部上に生化学物質のＤＮＡに特有な塩基配
列から構成された所望のＤＮＡ断片を固定し、 前記ＤＮＡ断片が固定された各々前記島部の共振周波数
を測定して前記島部各々の第１の測定周波数とし、 前記島部上に固定された前記ＤＮＡ断片を検体となるＤ
ＮＡを含んだ溶液中に所定時間接触させた後で乾燥し、
各々の前記島部の共振周波数を測定して前記島部各々の
第２の測定周波数とし、 前記第２の測定周波数と第１の測定周波数との差によ
り、前記検体となるＤＮＡの中より前記ＤＮＡ断片と同
じ塩基配列を検出することを特徴とするＤＮＡ検査方
法。7. A quartz substrate provided with a plurality of islands each separated at a predetermined interval, and a desired DNA fragment composed of a base sequence peculiar to a DNA of a biochemical substance is provided on each of the islands. And the resonance frequency of each of the islands to which the DNA fragment is fixed is measured to be the first measurement frequency of each of the islands, and the DNA fragment fixed on the island is used as a sample D
After contacting in a solution containing NA for a predetermined time, it is dried,
The resonance frequency of each of the islands is measured to obtain a second measurement frequency of each of the islands, and the difference between the second measurement frequency and the first measurement frequency is used to select the DNA from the DNA to be the sample. A method for testing DNA, which comprises detecting the same nucleotide sequence as a DNA fragment.
て、 前記島部は、前記水晶基板上に形成された凸部であるこ
とを特徴とするＤＮＡ検査方法。8. The DNA inspection method according to claim 7, wherein the island portion is a convex portion formed on the quartz substrate.
て、 前記島部は、前記水晶基板上に形成された凹部であるこ
とを特徴とするＤＮＡ検査方法。9. The DNA inspection method according to claim 7, wherein the island portion is a recess formed on the quartz substrate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002285845A JP4094922B2 (en) | 2002-01-28 | 2002-09-30 | DNA chip and DNA testing method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002018148 | 2002-01-28 | ||
| JP2002-18148 | 2002-01-28 | ||
| JP2002285845A JP4094922B2 (en) | 2002-01-28 | 2002-09-30 | DNA chip and DNA testing method |
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| JP4094922B2 JP4094922B2 (en) | 2008-06-04 |
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| JP2002285845A Expired - Fee Related JP4094922B2 (en) | 2002-01-28 | 2002-09-30 | DNA chip and DNA testing method |
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