JPH1144690A - Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using it - Google Patents
Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using itInfo
- Publication number
- JPH1144690A JPH1144690A JP19972297A JP19972297A JPH1144690A JP H1144690 A JPH1144690 A JP H1144690A JP 19972297 A JP19972297 A JP 19972297A JP 19972297 A JP19972297 A JP 19972297A JP H1144690 A JPH1144690 A JP H1144690A
- Authority
- JP
- Japan
- Prior art keywords
- sludge
- antibody
- bacillus
- bacillus bacteria
- detecting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、バチルス属細菌検
出用抗体およびこれを用いた汚泥中のバチルス属細菌の
検出法に関し、さらに詳しくは下水処理、工場排水処理
等に使用される生物的処理において、活性汚泥中に含ま
れる有用細菌であるグラム陽性細菌のバチルス属細菌を
迅速に検出し、その処理能力を迅速に把握することがで
きるバチルス属細菌検出用抗体およびこれを用いた汚泥
中のバチルス属細菌の検出法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antibody for detecting Bacillus bacteria and a method for detecting Bacillus bacteria in sludge using the same, and more particularly to a biological treatment used for sewage treatment, industrial wastewater treatment, and the like. In, the Bacillus genus bacteria that are useful bacteria contained in the activated sludge, which is a gram-positive bacterium, can be quickly detected, and its processing ability can be quickly grasped. The present invention relates to a method for detecting Bacillus bacteria.
【０００２】[0002]
【従来の技術】従来、活性汚泥中の微生物の菌種、菌数
についての同定または定量は、サンプリングした汚泥に
ついて種々の条件で培養試験を行うことによりなされて
いた。特にバチルス属菌体に関しては、その広い栄養要
求性から有効な選抜培地が存在しないため、コロニーを
単離し、その形態、グラム染色性等を調査した後、類似
した形のコロニーの数を調べ、その数をもって汚泥中の
バチルス属細菌数としてきた。2. Description of the Related Art Conventionally, the identification or quantification of the microorganism species and the number of microorganisms in activated sludge has been carried out by conducting a culture test on the sampled sludge under various conditions. In particular, for Bacillus sp., Since there is no effective selection medium due to its wide auxotrophy, colonies were isolated, their morphology, gram stainability, etc. were investigated, and then the number of colonies of similar shape was examined. The number was used as the number of Bacillus bacteria in the sludge.
【０００３】しかしながら、前記培養試験による方法で
は、最終的にはコロニーの形態によりバチルス属細菌数
を測定することになるため、コロニーがバチルス属特有
の形態を示す頃には、コロニーの直径が大きくなり、コ
ロニー同志が重なり合って正確な菌数の把握が不可能で
あることがしばしばであった。またこの方法は培養操作
が必要であるため、１〜３日間の培養期間が必要であ
る。また、この方法では菌種レベルでの計測は不可能で
あり、特に有機物分解能に優れたバチルス属の菌種を検
出しようとしても、平板上のコロニーでは区別をつける
ことは不可能であった。However, in the method based on the culture test, the number of Bacillus bacteria is ultimately measured based on the morphology of the colony. Therefore, by the time the colony shows a morphology unique to the genus Bacillus, the diameter of the colony increases. In many cases, accurate colony counts were impossible due to overlapping of colonies. In addition, since this method requires a culturing operation, a culturing period of 1 to 3 days is required. In addition, it is impossible to measure at the bacterial species level by this method. In particular, even if it is intended to detect a bacterial species of the genus Bacillus having excellent organic substance resolution, it is impossible to distinguish between colonies on a plate.
【０００４】[0004]
【発明が解決しようとする課題】本発明の課題は、上記
従来技術の問題点を解決し、汚泥中のバチルス属細菌を
迅速かつ簡便に検出、定量することができるバチルス属
細菌検出用抗体およびこれを用いた汚泥中のバチルス属
細菌の検出法を提供することにある。An object of the present invention is to solve the above-mentioned problems of the prior art, and to provide an antibody for detecting a Bacillus bacterium capable of rapidly and easily detecting and quantifying a Bacillus bacterium in sludge. An object of the present invention is to provide a method for detecting Bacillus bacteria in sludge using the same.
【課題を解決するための手段】本発明者らは、前記状況
を踏まえ、汚泥中のバチルス属細菌を迅速かつ簡便に検
出定量することを目標として鋭意研究した結果、バチル
ス属細菌に対して特異的に作用する抗体を用い、その反
応特性、特にその反応特性と相関する発色反応を指標と
してバチルス属細菌を検出し、定量することにより、所
期の目標を達成し得ることを見い出し、本発明を完成す
るに至った。Means for Solving the Problems Based on the above situation, the present inventors have conducted intensive studies with the aim of detecting and quantifying Bacillus bacteria in sludge quickly and easily. The present invention has been found to be able to achieve the intended target by detecting and quantifying Bacillus bacteria using an antibody that acts in a specific manner and using its reaction characteristics, particularly a color reaction correlated with the reaction characteristics, as an index. Was completed.
【０００５】すなわち、本発明で特許請求される発明は
以下のとおりである。 （１）バチルス属細菌の菌体外分泌物を免疫原として得
られたバチルス属細菌検出用抗体。 （２）前記バチルス属細菌検出用抗体を、汚泥中のバチ
ルス属細菌に特異的に反応させて汚泥中のバチルス属細
菌を検出することを特徴とする汚泥中のバチルス属細菌
の検出法。 （３）前記バチルス属細菌検出用抗体を、汚泥中のバチ
ルス属細菌に特異的に反応させ、該反応と相関する発色
反応を指標として汚泥中のバチルス属細菌の定量を行う
ことを特徴とする汚泥中のバチルス属細菌の検出法。That is, the invention claimed in the present invention is as follows. (1) An antibody for detecting a Bacillus bacterium obtained by using an extracellular secretion of a Bacillus bacterium as an immunogen. (2) A method for detecting Bacillus bacteria in sludge, wherein the Bacillus bacteria-detecting antibody is specifically reacted with the Bacillus bacteria in sludge to detect Bacillus bacteria in sludge. (3) The antibody for Bacillus genus detection is caused to react specifically with the Bacillus bacterium in sludge, and the Bacillus bacterium in the sludge is quantified using a color reaction correlated with the reaction as an index. Method for detecting Bacillus bacteria in sludge.
【０００６】[0006]
【発明の実施の形態】本発明におけるバチルス属細菌検
出用抗体は、バチルス属細菌の菌体外分泌物を免疫原と
して用いることにより得られる。このバチルス属細菌の
菌体外分泌物は、公知の方法でバチルス属細菌を培養
し、得られた培養液を遠心分離等により菌体分画と菌体
分泌物分画を分離することにより得ることができる。バ
チルス属細菌の菌体外分泌物を免疫動物血中に接種する
と、該動物血中でバチルス属細菌に特異的に反応する抗
体が迅速に産生され、抗体価の高い抗血清が得られる。
抗体価の高い抗血清が得られれば、この抗血清をバチル
ス属細菌検出用抗体として使用することができる。また
抗体の精製が必要な場合においても、精製操作が容易に
なる。BEST MODE FOR CARRYING OUT THE INVENTION The antibody for detecting a Bacillus bacterium according to the present invention can be obtained by using an extracellular secretion of a Bacillus bacterium as an immunogen. The extracellular secretion of the Bacillus bacterium is obtained by culturing the Bacillus bacterium by a known method, and separating the obtained culture solution into a cell fraction and a cell secretion fraction by centrifugation or the like. Can be. When an extracellular secretion of a Bacillus bacterium is inoculated into the blood of an immunized animal, an antibody specifically reacting with the Bacillus bacterium is rapidly produced in the blood of the animal, and an antiserum having a high antibody titer is obtained.
If an antiserum having a high antibody titer can be obtained, this antiserum can be used as an antibody for detecting Bacillus bacteria. Further, even when antibody purification is required, the purification operation is facilitated.
【０００７】上記抗血清（抗体）は、９６穴ウェルプレ
ート、スチレンボール等に容易に固相化することが可能
であり、また固相化した後、ＢＳＡ等の一般的なブロッ
キング剤によりブロッキングすることにより、これを冷
蔵で保存することが可能となる。本発明において、汚泥
中のバチルス属細菌の検出は、前記抗体を汚泥中のバチ
ルス属細菌（抗原）に特異的に反応させ、その特異的な
反応特性を検知することにより行われる。この反応を検
知する方法としては、酸素や蛍光色素により標識された
二次抗体を用いる方法、抗体と抗原（バチルス属細菌）
との結合により生じた凝集を検出する方法等が挙げられ
る。The above-mentioned antiserum (antibody) can be easily immobilized on a 96-well plate, a styrene ball or the like. After immobilization, the antiserum is blocked with a general blocking agent such as BSA. This makes it possible to store it refrigerated. In the present invention, the detection of Bacillus bacteria in sludge is carried out by specifically reacting the antibody with Bacillus bacteria (antigen) in sludge and detecting the specific reaction characteristics. As a method of detecting this reaction, a method using a secondary antibody labeled with oxygen or a fluorescent dye, an antibody and an antigen (Bacillus bacterium)
And a method for detecting agglutination caused by the binding to the enzyme.
【０００８】前者の方法は、抗体とスチレンやガラスが
結合する性質を利用して行われる。例えば、あらかじめ
スチレン製の９６孔ウェルプレート、ボール、ガラスビ
ーズ等の表面にアルカリ条件下で抗体や血清を作用させ
て固定化し、その後、スチレンやガラス表面の抗体の固
定化されていない部分を、抗体を含まない蛋白質（ＢＳ
Ａ）で不活性化（ブロッキング）し、次いで該９６穴ウ
ェルプレート等に、抗原を含んだ溶液を作用させて抗体
に抗原を吸着させた後、標識した二次抗体を作用させて
抗原抗体結合物を検出する。[0008] The former method is carried out by utilizing the property of binding an antibody to styrene or glass. For example, 96-well plate made of styrene, balls, glass beads, etc., are immobilized by applying an antibody or serum under alkaline conditions in advance, and then, the non-immobilized portion of the styrene or glass surface is fixed to the antibody. Antibody-free protein (BS
A) is inactivated (blocked) in step A), and then a solution containing the antigen is allowed to act on the 96-well plate or the like to allow the antigen to be adsorbed to the antibody. Detect objects.
【０００９】二次抗体には、通常、一次抗体を作製した
動物種の血清に結合する抗体や、抗原に対して結合する
抗体を標識したものなどが用いられ、二次抗体の標識と
して酵素を用いる場合の方法はＥＬＩＳＡ法（酵素標識
固槽免疫測定法）として知られている。ＥＬＳＩＡ法に
用いる二次抗体には、通常、ペルオキシダーゼ、アルカ
リフォスファターゼ等の基質に対する反応時に共役して
色素を変化させる酵素が付加されており、抗原抗体結合
物に二次抗体が作用した反応系に対して色素を含んだ基
質液を添加して基質液中の色素が変色すれば、抗原（バ
チルス属細菌）の存在が確認される。変色の有無まはた
変色の程度は、吸光度の測定により行われる。また菌体
数と変色量の関係をあらかじめ調べて検量線を作成して
おくことにより、菌体の定量を迅速かつ簡便に行うこと
ができる。[0009] As the secondary antibody, an antibody that binds to the serum of the animal species from which the primary antibody was prepared, or an antibody that binds to an antigen is usually used, and an enzyme is used as the secondary antibody label. The method when used is known as ELISA (enzyme-labeled solid-tank immunoassay). The secondary antibody used in the ELISA method is usually added with an enzyme that conjugates and changes the dye at the time of reaction with a substrate such as peroxidase or alkaline phosphatase. On the other hand, if the color of the dye in the substrate solution is changed by adding the substrate solution containing the dye, the presence of the antigen (Bacillus bacterium) is confirmed. The presence or absence of discoloration or the degree of discoloration is determined by measuring absorbance. In addition, by determining the relationship between the number of cells and the amount of discoloration in advance and preparing a calibration curve, quantification of the cells can be performed quickly and easily.
【００１０】[0010]
【実施例】以下、本発明を実施例により詳しく説明する
が、本発明はこれらに限定されるものではない。なお、
例中の％は特に限定しない限り重量％を意味する。 実施例１ 表１に示した菌体培養培地に、活性汚泥より単離したＢ
ａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｅｎｓｉｓを接種し、
３７℃、好気的条件下で培養した。この培養液を遠心分
離にかけ、沈降したペレットに水を加えて懸濁させ、こ
れを粗調製液とした。この粗調製液を再び遠心分離し、
上澄みと沈殿とに分離した。この操作を３回繰り返し、
最終的に得られた沈殿物を菌体分画、粗調製液の上澄み
を菌体分泌物分画とした。また粗調製液に対してトリク
ロロ酢酸を１％の濃度になるように加え、よく懸濁した
後、遠心分離し、さらに沈殿物に１％トリクロロ酢酸溶
液を加えて懸濁させた後、再び遠心分離した。これらの
操作で得られた上澄みにメタノールを加え、沈殿した成
分を水に溶解させてＴＣＡ分画とした。The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the invention thereto. In addition,
Unless otherwise specified,% in the examples means% by weight. Example 1 B cells isolated from activated sludge were added to the cell culture medium shown in Table 1.
acillus thuringensis ,
The cells were cultured at 37 ° C under aerobic conditions. This culture solution was centrifuged, and water was added to the precipitated pellet to suspend it, which was used as a crude preparation. This crude preparation is centrifuged again,
It was separated into a supernatant and a precipitate. Repeat this operation three times,
The finally obtained precipitate was used as a cell fraction, and the supernatant of the crude preparation was used as a cell secretion fraction. Trichloroacetic acid was added to the crude preparation so as to have a concentration of 1%, suspended well, centrifuged, and the precipitate was suspended by adding a 1% trichloroacetic acid solution, and then centrifuged again. separated. Methanol was added to the supernatant obtained by these operations, and the precipitated component was dissolved in water to obtain a TCA fraction.
【００１１】得られた菌体分画、菌体分泌物分画および
ＴＣＡ分画をそれぞれ凍結乾燥させ、得られた粉末をア
ジュバントにし、これを白色兎に対して１０日に１回の
割合で接種し、血液を採取してそのときの抗体価をＭＡ
Ｐ抗原に対するＥＬＩＳＡ法により測定した。その結果
を表２に示す。なお、抗体価は４１５ｎｍ、ＯＤ≒０．
２の時の希釈倍率で示した。[0011] The obtained cell fraction, cell secretion fraction and TCA fraction are each freeze-dried, and the resulting powder is adjuvanted to white rabbits once every 10 days. After inoculation, blood was collected and the antibody titer was
It was measured by an ELISA method for the P antigen. Table 2 shows the results. The antibody titer was 415 nm and OD ≒ 0.
The dilution ratio at the time of 2 was shown.
【００１２】[0012]
【表１】 [Table 1]
【００１３】[0013]
【表２】 表２の結果から、菌体外分泌物を動物の血中に接種する
ことにより、効率的に高い抗体価の抗体が免疫動物血中
に生産されることが確認された。[Table 2] From the results in Table 2, it was confirmed that inoculation of the extracellular secretions into the blood of the animal efficiently produced an antibody having a high antibody titer in the blood of the immunized animal.
【００１４】実施例２Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｅｎｓｉｓ 、Ｅｓｃ
ｈｅｒｉｃｈｉａ Ｃｏｌｉ（ＡＴＣＣ ２５９２２）
およびＰａｒａｃｏｃｃｕｓ ｄｅｎｉｔｒｉｆｉｃｏ
ｉｎｓ（ＩＦＯ １３３０１）の菌株を表１に示した菌
体培養培地にそれぞれ接種し、３７℃、好気的条件下で
培養した。得られた培養液をリン酸緩衝液（ＰＢＳ）で
１０8 個／ｍｌになるように希釈し、実施例１で得られ
た抗体を９６穴ウェルプレートに固相化し、ＥＬＩＳＡ
法により、各菌体の抗体に対する特異性を調べ、その結
果を表３に示した。吸光度測定は４１５ｎｍで行い、菌
液を添加しないブランクの吸光度より高い場合には＋で
示し、低いかほとんど同じ場合には−で示した。Example 2 Bacillus thuringensis , Esc
herichia Coli (ATCC 25922)
And Paracoccus denitrifico
The ins (IFO 13301) strain was inoculated into the cell culture medium shown in Table 1 and cultured at 37 ° C under aerobic conditions. The obtained culture solution was diluted with phosphate buffer (PBS) to 10 8 cells / ml, the antibody obtained in Example 1 was immobilized on a 96-well plate, and ELISA was performed.
The specificity of each cell to the antibody was examined by the method, and the results are shown in Table 3. The absorbance was measured at 415 nm, and was indicated by + when the absorbance was higher than that of the blank without addition of the bacterial solution, and by-when the absorbance was low or almost the same.
【００１５】[0015]
【表３】 表３の結果から、実施例１で得られた抗体は、Ｂａｃｉ
ｌｌｕｓ ｔｈｕｒｉｎｇｅｎｓｉｓに対して特異的に
反応することが確認された。[Table 3] From the results of Table 3, the antibody obtained in Example 1, Baci
It was confirmed that the antibody specifically reacted with L. thuringensis .
【００１６】実施例３ 実施例１で培養したＢａｃｉｌｌｕｓ ｔｈｕｒｉｎｇ
ｅｎｓｉｓを１．２５×１０5 個／ｍｌ〜６×１０6 個
／ｍｌの範囲で希釈し、実施例１で得られた抗体を９６
穴ウェルプレートに固相化し、ＥＬＩＳＡ法により菌体
数に対する吸光度を測定し、検量線を作成した。得られ
た検量線を図１に示したが、縦軸を対数とした片対数に
よる最小２乗法による相関をとると、吸光度＝−３．３
５＋０．６６７×ｌｏｇ（細胞数）という式が得られ
た。以上の結果から、汚泥中のＢａｃｉｌｌｕｓ ｔｈ
ｕｒｉｎｇｅｎｓｉｓを同様の操作で本発明の抗体と反
応させ、ＥＬＩＳＡ法によりその吸光度を測定すること
により、従来のように培養操作を経ずに容易に菌体数の
定量ができることがわかった。Example 3 Bacillus thuring cultured in Example 1
ensis diluted in the range of 1.25 × 10 5 cells / ml~6 × 10 6 cells / ml, and the antibody obtained in Example 1 96
The solid phase was immobilized on a well plate, and the absorbance with respect to the number of cells was measured by an ELISA method to prepare a calibration curve. The obtained calibration curve is shown in FIG. 1. When the correlation was obtained by the least square method using semilogarithm with the logarithm on the vertical axis, the absorbance was −3.3.
The formula 5 + 0.667 × log (number of cells) was obtained. From the above results, Bacillus th in sludge
uringensis was reacted with the antibody of the present invention in the same manner, and the absorbance was measured by ELISA. Thus, it was found that the number of cells could be easily quantified without a conventional culture operation.
【００１７】実施例４ 活性汚泥３ｇ−ＦＷ（湿重量）に、ＢＯＤ５００ｐｐｍ
の人口汚水を１リットル添加し、さらにＢａｃｉｌｌｕ
ｓ ｔｈｕｒｉｎｇｅｎｓｉｓを接種して通気しながら
４日間の培養を行った。このときに用いた培養培地の成
分を表４に示した。次いで、実施例１で得られた抗体を
９６穴ウエルプレートに固定化し、ＢＳＡでブロッキン
グした後、固定化した抗体に上記の培養で得られた培養
液を添加して反応させ、ＥＬＩＳＡ法によりＢａｃｉｌ
ｌｕｓ ｔｈｕｒｉｎｇｅｎｓｉｓの菌体数を、実施例
３で得られた検量線に基づいて定量したところ１．２×
１０ 6 個／ｍｌであった。Example 4 Activated sludge 3 g-FW (wet weight), BOD 500 ppm
1 liter of artificial wastewaterBacillu
s thuringensisInoculate and ventilate
Culture was performed for 4 days. The culture medium used at this time
The minutes are shown in Table 4. Next, the antibody obtained in Example 1 was
Immobilize on 96-well plate and block with BSA
After the culture, the immobilized antibody
The reaction is performed by adding the liquid, and the ELISA method is used.Bacil
rus thuringensisExample 1
As a result of quantification based on the calibration curve obtained in 3, 1.2 ×
10 6Pcs / ml.
【００１８】[0018]
【表４】 [Table 4]
【００１９】実施例５ 実施例４において、Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇ
ｅｎｓｉｓの培養を通気しないで行った以外は、実施例
４と同様にしてＢａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｅｎ
ｓｉｓの培養と菌体数の定量を行った。しかし、培養条
件が嫌気状態であったため、絶対好気性菌であるＢａｃ
ｉｌｌｕｓ ｔｈｕｒｉｎｇｅｎｓｉｓが死滅し、菌体
は検出されなかった。Example 5 In Example 4, Bacillus thuring
Bacillus thuringen in the same manner as in Example 4 except that the culture of B. ensis was performed without aeration.
The sis was cultured and the number of cells was quantified. However, since the culture conditions were anaerobic, Bac , an absolute aerobic bacterium, was used.
illus thuringensis died, and no bacterial cells were detected.
【００２０】実施例６ 実施例４において、Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇ
ｅｎｓｉｓを接種せず通気しないで培養を行った以外は
実施例４と同様にしてＢａｃｉｌｌｕｓ ｔｈｕｒｉｎ
ｇｅｎｓｉｓの培養と菌体数の定量を行ったが、菌体は
検出されなかった。Example 6 In Example 4, Bacillus thuring
Bacillus Thurin except that the culture was performed without aeration without inoculated ensis in the same manner as in Example 4
Gengen was cultured and the number of cells was determined, but no cells were detected.
【００２１】[0021]
【発明の効果】本発明におけるバチスル属細菌検出用抗
体は、バチルス属細菌の菌体外分泌物を免疫原として容
易に得ることができ、バチルス属細菌に特異的に反応す
るため、その反応特性を利用して汚泥中のバチルス属細
菌の検出を容易に行うことができる。またその反応特性
と相関する発色反応を指標とすることにより、予め作成
した検量線に基づいて迅速かつ簡便に汚泥中のバチルス
属細菌の定量を行うことができる。The antibody for detecting Bacillus genus bacteria according to the present invention can easily obtain an extracellular secretion of Bacillus bacterium as an immunogen and reacts specifically with Bacillus bacterium. Bacillus bacteria in sludge can be easily detected by utilizing the method. In addition, by using a coloring reaction correlated with the reaction characteristics as an index, it is possible to quickly and easily quantify Bacillus bacteria in sludge based on a previously prepared calibration curve.
【図１】実施例３で得られた菌体濃度と吸光度にかかわ
る検量線を示した図。FIG. 1 is a diagram showing a calibration curve relating to the bacterial cell concentration and absorbance obtained in Example 3.
Claims (3)
として得られたバチルス属細菌検出用抗体。1. An antibody for detecting a Bacillus bacterium obtained by using an extracellular secretion of a Bacillus bacterium as an immunogen.
体を、汚泥中のバチルス属細菌に特異的に反応させて汚
泥中のバチルス属細菌を検出することを特徴とする汚泥
中のバチルス属細菌の検出法。2. The Bacillus genus in sludge, wherein the Bacillus bacterium detection antibody according to claim 1 is specifically reacted with the Bacillus bacterium in the sludge to detect the Bacillus bacterium in the sludge. Bacteria detection method.
体を、汚泥中のバチルス属細菌に特異的に反応させ、該
反応と相関する発色反応を指標として汚泥中のバチルス
属細菌の定量を行うことを特徴とする汚泥中のバチルス
属細菌の検出法。3. The antibody for Bacillus genus detection according to claim 1 is caused to react specifically with Bacillus bacterium in sludge, and quantification of Bacillus bacterium in sludge is performed using a color reaction correlated with the reaction as an index. A method for detecting Bacillus bacteria in sludge.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19972297A JPH1144690A (en) | 1997-07-25 | 1997-07-25 | Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using it |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP19972297A JPH1144690A (en) | 1997-07-25 | 1997-07-25 | Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using it |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH1144690A true JPH1144690A (en) | 1999-02-16 |
Family
ID=16412531
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP19972297A Pending JPH1144690A (en) | 1997-07-25 | 1997-07-25 | Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using it |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH1144690A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010082590A (en) * | 2008-10-01 | 2010-04-15 | Sumiju Kankyo Engineering Kk | Simple measuring method for bacillus bacteria |
| JP2011512861A (en) * | 2008-03-14 | 2011-04-28 | ビオメリュー | A method for real-time detection of microorganisms in liquid media by agglutination |
-
1997
- 1997-07-25 JP JP19972297A patent/JPH1144690A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011512861A (en) * | 2008-03-14 | 2011-04-28 | ビオメリュー | A method for real-time detection of microorganisms in liquid media by agglutination |
| JP2010082590A (en) * | 2008-10-01 | 2010-04-15 | Sumiju Kankyo Engineering Kk | Simple measuring method for bacillus bacteria |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4497900A (en) | Immunoassay for Neisseria gonorrhoeae antigens | |
| US4497899A (en) | Immunoassay for Chlamydia trachomatis antigens | |
| CN101971032B (en) | Method for the real-time detection of microorganisms in a liquid culture medium by agglutination | |
| US5750357A (en) | Method of rapid analyte detection | |
| Monteiro et al. | Evaluation of performances of three DNA enzyme immunoassays for detection of Helicobacter pylori PCR products from biopsy specimens | |
| US5126244A (en) | Determination of antigens | |
| MY103181A (en) | Detection of cell membrane protein. | |
| JPH1144690A (en) | Antibody for detecting bacillus bacteria and method for detecting bacillus bacteria in sludge using it | |
| US8765399B2 (en) | Cultures and protocols for diagnosis of toxigenic Clostridium difficile | |
| AU2007310738B2 (en) | Method for in vitro diagnosis of PVL-producing Staphylococcus aureus | |
| Ataee et al. | Staphylococcal Enterotoxin A Detection from Rheumatoid Arthritis Patients’ Blood and Synovial Fluid | |
| CA2319575C (en) | Method for the determination of prosthetic infections | |
| Thorne et al. | Enzymatically labelled nucleic acid (NA) probe assays for detection of Campylobacter spp. in human faecal specimens and in culture | |
| Zambardi et al. | European multicentre evaluation of a commercial system for identification of methicillin-resistant Staphylococcus aureus | |
| Zueter et al. | Current protocols in laboratory diagnosis, genotyping, and treatment of Burkholderia pseudomallei | |
| JP4035738B2 (en) | Novel alkaline phosphatase | |
| KR101094974B1 (en) | Novel Biomarkers For the Serodiagnosis of Crohn?s disease | |
| Pietrzak et al. | Sandwich-dot immunobinding assay (sandwich-DIA), a new immunological method for the detection of diphtheria toxin | |
| Aleixo et al. | A fluorescent enzyme immunoassay for Salmonella detection | |
| Young et al. | Utility of monoclonal antibody coagglutination to identify Neisseria gonorrhoeae. | |
| Akimenko et al. | A rapid method for detection of Clostridium thermocellum by field-effect transistor-based immunodetection | |
| Gürtürk et al. | Detection of Campylobacter Antibodies in Sheep Sera by a Dot‐ELISA Using Acid Extracts from C. fetus ssp. fetus and C. jejuni Strains and Comparison with a Complement Fixation Test | |
| CZ280282B6 (en) | Enzymatic determination method of antibiotics with beta-lactam core | |
| Denis et al. | A latex agglutination technique for rapid, direct identification of pneumococci in blood cultures | |
| Robinson et al. | Evaluation of the RIM system and Gono Gen test for identification of Neisseria gonorrhoeae from clinical specimens |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Effective date: 20040609 Free format text: JAPANESE INTERMEDIATE CODE: A621 |
|
| A977 | Report on retrieval |
Effective date: 20051102 Free format text: JAPANESE INTERMEDIATE CODE: A971007 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20051108 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20060314 |