CN1673745A - Process for enzyme-linked immunosorbent ELISA synchronous fast assay method for multiple immune indexes - Google Patents
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Abstract
The present invention provides one enzyme-linked immunosobant assay (ELISA) method for synchronous fast detection of several immune indexes. The method includes the following steps: coating two or more kinds of specific antibodies or antigens in the same enzyme-linked plate holes and drying in the shade at 4 deg.c after adsorption; extracting patient's sample and adding into the plate holes; adding corresponding known antibodies or antigens into the plate holes and washing; adding one kind of substrate, developing, comparing with contrast and washing; and repeating with one other kind of substrate until finishing the detection. The present invention can detect several immune indexes in once operation, and is fast, simple and low in cost.
Description
Technical field
The present invention relates to medical microbial and Medical Immunology detection technique, relate in particular to a kind of method of measuring immune indexes by enzyme linked immunosorbent assay (ELISA), more specifically say, relate to a kind of method by the enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes
Background technology
Enzyme linked immunosorbent assay (ELISA) is a class important technology of clinical both at home and abroad and experimental study, relate to medical science, biology, science of heredity, many science such as immunity, its ultimate principle is to utilize antigen, the specific reaction of antibody, antibody or antigen with known antigen or antibody test the unknown, particularly extensive aspect the observation of the diagnosis of disease and curative effect and preventive medicine, as medically in order to diagnosis and differential diagnosis hepatitis virus (first, second, third, fourth, penta, Hepatitis G Virus), AIDS diagnosis and other bacterium, Chlamydia, the diagnosis of protozoan infection.
Make a definite diagnosis with enzyme connection Elisa method and detect the method that pathogen is the WHO regulation, this method is generally competition law or sandwich method.
Existing double antibody sandwich method is that specific antibody (monoclonal, polyclone) is formed insolubilized antibody with the Elisa plate, utilizes the same antibody (monoclonal, polyclone) of enzyme labeling to combine the method that detects a kind of antigen: basic operational steps is as follows:
1, antibody sandwich.Specific antibody is combined with carrier, and behind the formation insolubilized antibody, flush away is binding antibody and impurity not, and 4 ℃ dry in the shade.
2, add the unknown antigen sample: allow sample antigen and insolubilized antibody fully react, afterwards flush away bound substances not.
3, add enzyme labelled antibody, allow other free determinants of antigen combine (this law is unsuitable for detecting haptens, monovalent antigen) in known enzyme labelled antibody and the solid-phase complex.
4, detect: the enzyme in the corresponding enzymic-labelled antibody adds chromogenic substrate, and compares with control group, according to the depth of substrate colour developing, judges to have or not antigen.
Existing competition law can be used for measuring antigen, also can be used for measuring antibody, but is used for measuring micromolecule antigen more, and basic operational steps is as follows:
1, antibody sandwich: specific antibody is combined with carrier, behind the formation insolubilized antibody, flush away not binding antibody and impurity.
2, add not key sample of patient;
3, add enzyme target known antigens, key sample antigen does not combine with insolubilized antibody competitively with known antigens through enzyme labeling, and flush away is bound substances not.
4, detect: the enzyme in the corresponding enzyme-labelled antigen adds chromogenic substrate, and compares with control group, and the depth that develops the color according to substrate has or not, and judges to have or not antigen.
In the above-mentioned Elisa method, for not judging whether contain pathogen (antigen) in the key sample, should prepare control group simultaneously, in the compound method of competition law control group, only save to bag and add the not step of key sample in by good elisa plate, all the other steps are identical.In the compound method of sandwich method control group, with step 2: add the step of unknown antigen sample, become to bag and add known antigen in by good enzyme linked plate holes, all the other steps are identical.When detecting, then in control group, add the detection substrate identical simultaneously with test sample, by the difference of the shade between control group and test sample, can judge in the not key sample to be detected whether contain pathogen.
More than can measure the OD value by microplate reader after the colour developing of two kinds of methods, perhaps by naked eyes according to the direct judged result of the depth of color.
Existing above-mentioned sandwich method and competition law once can only be measured a kind of pathogen, as detecting multiple pathogen, need repeatedly prepare respectively, antigen or antibody are checked respectively, and complex steps, detection time is long, wastes time and energy, and detects the cost height.
Summary of the invention
The present invention has overcome existing enzyme linked immunosorbent assay (ELISA) once can only detect a kind of pathogen, when detecting multiple pathogen, the shortcoming of can only operate respectively, prepare, checking, a kind of method of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes is provided, this method can be prepared by single job, the synchronous detection multiple immune indexes, and detection speed is fast, save human and material resources, reduce and detect cost.
For realizing purpose of the present invention, the following technical proposals that the present invention adopts.
The invention discloses a kind of method of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes, i.e. double antibody method comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) specific antibody with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in (antigen pathogen) sample adding enzyme linked plate holes of patient;
(3) kind and the quantity of specific antibody in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antibodies of quantity, wash plate then;
(2) preparation control group
(1) method is identical with preparation test sample step (1);
(2) kind and the quantity of specific antibody in the corresponding step (1) join the known antigens of identical kind and quantity in the enzyme linked plate holes simultaneously,
(3) method is identical with preparation test sample step (3), and the kind and the quantity of specific antibody in the promptly corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antibodies of quantity, fully wash plate after the reaction;
(3) control test
A kind of enzyme in corresponding test sample and the control group step (3) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
The present invention discloses the method for another kind of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes, i.e. competition law comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) specific antibody with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in (antigen pathogen) sample adding enzyme linked plate holes of patient, and simultaneously
(3) kind and the quantity of specific antibody in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antigens of quantity, fully wash plate after the reaction;
(2) preparation control group
(1) method is identical with the step (1) of preparation test sample;
(2) method is identical with the step (3) of preparation test sample, and the kind and the quantity of specific antibody in the promptly corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antigens of quantity, fully wash plate after the reaction;
(3) control test
A kind of enzyme in corresponding test sample step (3) and the control group step (2) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
The present invention further discloses the method for another kind of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes, comprise the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) the antigen pathogen with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in patient's the sample adding enzyme linked plate holes;
(3) kind and the quantity of antigen pathogen in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known specific antibody of quantity, fully wash plate after the reaction;
(2) preparation control group
(1) method is identical with the step (1) of preparation test sample;
(2) method is identical with the step (3) of preparation test sample;
(3) control test
A kind of enzyme in corresponding test sample step (3) and the control group step (2) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
In the technique scheme, in first and second kinds of methods, the specific antibody that is about to two or more in the wrapper sheet process is coated in the enzyme linked plate holes, as rotavirus specific antibody and adenovirus specific antibody are coated in the enzyme linked plate holes, after the absorption, 4 ℃ dry in the shade; The sample that extracts patient afterwards adds in the enzyme linked plate holes.
When the sandwich method among the employing enzyme linked immunological absorption ELISA, behind the not key sample that adds patient, in the corresponding step (1) value volume and range of product of specific antibody with identical value volume and range of product, join in the enzyme linked plate holes with the known antibody of different enzyme labelings, as joining in the enzyme linked plate holes through the rotavirus antibody and the adenovirus antibody of different enzyme labelings, fully after the reaction, flush away is bound substances not.
When adopting competition law, behind the not key sample that adds patient, in the corresponding step (1) quantity of specific antibody with identical value volume and range of product, join in the enzyme linked plate holes with the known antigen of different enzyme labelings, as joining in the enzyme linked plate holes through the wheel virus antigen and the adenovirus antigen of different enzyme labelings, key sample antigen does not combine with insolubilized antibody competitively with known antigens through enzyme labeling, fully after the reaction, flush away is bound substances not.
For not judging whether contain pathogen in the key sample, need the preparation control group, in the sandwich method, the compound method of control group is: the specific antibody of two or more that will be identical with test sample is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade; The kind and the quantity of corresponding specific antibody join the known antigens of equal kind and quantity in the enzyme linked plate holes; The kind and the quantity of corresponding specific antibody will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antibodies of quantity, fully wash plate after the reaction.
In the competition law, the compound method of control group is: the specific antibody of two or more that will be identical with test sample is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade; The kind and the quantity of corresponding specific antibody will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antigens of quantity, fully wash plate after the reaction.
In above-mentioned two kinds of methods, after the preparation of test sample and control group is finished, a kind of enzyme in corresponding test sample and the control group plurality of enzymes adds a kind of substrate simultaneously earlier, colour developing is washed plate after relatively detecting, corresponding afterwards another kind of enzyme adds another kind of substrate, colour developing is washed plate after relatively detecting, and finishes to detecting successively.
As in competition law, do not add not key sample in the control group, after adding known different chromogenic enzyme substrate, control group and mensuration group relatively do not add in the control wells of sample, and color is the darkest, detect the poor of hole and control wells color depth, reflected antigen amount in the sample, that is to say that detection hole color is light more, illustrated that antigen in the sample (pathogen) content is many more.According to the colour developing depth and color difference, judge that unknown pathogen exists and content in the sample.
In the third method, in wrapping by process, two or more antigen pathogen is coated in the same enzyme linked plate holes, as rubella virus antigen and herpesviral is antigen coated in elisa plate, after adding patient's serum specimen, the kind and the quantity of corresponding antigen pathogen, will be with the identical kind of enzyme labeling of the same race and the known specific antibody of quantity do not join in the enzyme linked plate holes, fully wash plate after the reaction, after adding different enzyme labeling substrates, cause different chromogenic reactions, and relatively can detect patient specimen with control group.
In the technique scheme, during control group,, can corresponding test sample realize repeatedly control test, therefore save trouble, laborsaving, shorten detection time and cost only by a process for preparation in preparation.
More than can measure the OD value by microplate reader after the colour developing of three kinds of methods, perhaps by naked eyes according to the having or not of color, the direct judged result of the depth.
And with three kinds or four strain specific antibodies or antigen coated in enzyme linked plate holes, corresponding afterwards three kinds or four kinds of known antibody or antigens through enzyme labeling that add add three kinds or four kinds of substrates successively, can realize two or more pathogen are detected.
Among the present invention, microwell plate can be Polyvinylchloride microwell plate or polystyrene micropore plate, preferably adopts polystyrene micropore plate.
In the above-mentioned detection step, with identical in the prior art, each step is provided with washes the plate step.
Only need to do once experiment among the present invention, then can record two or more pathogen, manpower and materials have been saved greatly, reduced cost, reach the purpose of quick diagnosis, this method is not only valuable to the diagnosis of medical microbial cause of disease, and epidemiology survey, screening, clinical diagnosis are had crucial meaning.
Embodiment
Embodiment 1
The method of enzyme-linked immunosorbent ELISA synchronous fast assay rotavirus and adenovirus pathogen (sandwich method) comprises the following order step: preparation test sample and control group:
(1) preparation test sample:
(1) rotavirus specific antibody and adenovirus specific antibody are coated in the same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) (antigen) sample of extraction patient adds in the enzyme linked plate holes of test sample;
(3) will join in the enzyme linked plate holes of test sample with the rotavirus antibody of HRP (horseradish peroxidase) mark and the adenovirus antibody of alkali phosphatase enzyme mark, fully wash plate after the reaction.
(2) preparation control group
(1) rotavirus specific antibody and adenovirus specific antibody are coated in the same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) wheel virus antigen and adenovirus antigen are added in the above-mentioned enzyme linked plate holes;
(3) will join in the enzyme linked plate holes with the rotavirus antibody of HRP (horseradish peroxidase) mark and the adenovirus antibody of alkali phosphatase enzyme mark, fully wash plate after the reaction.
(3) control test
The substrate (PNPP) that in test sample and control group, adds detection of alkaline phosphatase simultaneously, fully after the reaction, test sample shows identical yellow with control group, does not then prove and contains the adenovirus pathogen in the key sample; Fully after the reaction, test sample is colourless, the control sample displaing yellow, does not prove not contain the adenovirus pathogen in the key sample.Fully clean enzyme linked plate holes, substrate is cleaned up (as colour developing, being about to enzyme linked plate holes is washed till colourless), in test sample and control group, add the substrate (TMB) that detects HRP (horseradish peroxidase) afterwards, test sample all shows identical avy blue with control group, does not then prove and contains rotavirus in the key sample; Fully after the reaction, test sample is colourless, the control sample displaing yellow, does not prove not contain the rotavirus pathogen, cessation reaction in the key sample.
Embodiment 2
The method of enzyme-linked immunosorbent ELISA synchronous fast assay rotavirus and adenovirus pathogen (competition law) comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) rotavirus specific antibody and adenovirus specific antibody are coated in the same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) extraction patient's sample (antigen) adds in the enzyme linked plate holes of test sample;
(3) will join simultaneously in the enzyme linked plate holes of test sample with the rotavirus of HRP (horseradish peroxidase) mark and the adenovirus antigen of alkali phosphatase enzyme mark, fully wash plate after the reaction;
(2) preparation control group
(1) rotavirus specific antibody and adenovirus specific antibody are coated in the same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) will join simultaneously in the enzyme linked plate holes of control group with the wheel virus antigen of HRP (horseradish peroxidase) mark and the adenovirus antigen of alkali phosphatase enzyme mark, fully wash plate after the reaction;
(3) control test
The substrate (PNPP) that in test sample and control group, adds detection of alkaline phosphatase, fully after the reaction, test sample shows identical yellow with control sample, and then not proving does not have adenovirus in the key sample; Fully after the reaction, test sample is colourless, the control sample displaing yellow, does not prove to contain the adenovirus pathogen in the key sample.Fully clean enzyme linked plate holes, substrate is cleaned up (as colour developing, being about to enzyme linked plate holes is washed till colourless), in test sample and control group, add the substrate (TMB) that detects HRP (horseradish peroxidase) afterwards, test sample shows identical blue look with control sample, and then not proving does not have rotavirus in the key sample; Fully after the reaction, test sample is colourless, and control sample shows blue look, does not prove and contains rotavirus in the key sample.Cessation reaction.
Embodiment 3
The method of enzyme-linked immunosorbent ELISA synchronous fast assay rubella virus and herpesviral comprises the following order step: preparation test sample and control group:
(1) preparation test sample:
(1) rubella virus antigen and herpesviral is antigen coated in same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) extract in patient's the enzyme linked plate holes of sample (specific antibody) as adding test sample such as serum;
(3) the rubella virus antibody of HRP mark and the herpesviral antibody of alkali phosphatase enzyme mark are joined in the enzyme linked plate holes of test sample, fully wash plate after the reaction.
(2) preparation control group
(1) rubella virus antigen and herpesviral is antigen coated in same polystyrene enzyme linked plate holes, 4 ℃ dry in the shade;
(2) the rubella virus antibody of HRP mark and the herpesviral antibody of alkali phosphatase enzyme mark are joined in the enzyme linked plate holes of test sample, fully wash plate after the reaction.
(3) control test
The substrate (PNPP) that in test sample and control group, adds detection of alkaline phosphatase simultaneously, fully after the reaction, test sample shows identical yellow with control group, does not then prove and does not contain the rubella virus pathogen antigen in the key sample; Fully after the reaction, test sample is colourless, the control sample displaing yellow, does not prove to contain the rubella virus pathogen antigen in the key sample.Fully clean enzyme linked plate holes, substrate is cleaned up (as colour developing, being about to enzyme linked plate holes is washed till colourless), in test sample and control group, add the substrate (TMB) that detects HRP (horseradish peroxidase) afterwards, test sample all shows identical avy blue with control group, does not then prove and does not contain herpesviral antibody in the key sample; Fully after the reaction, test sample is colourless, control sample shows blue look, does not prove and contains herpesviral antibody in the key sample.Cessation reaction.
In the embodiment 1,2,3,,, realize the check of multiple index in the key sample not by being coated with multiple known antibodies or antigen in same enzyme linked plate holes by one group of control group corresponding with test sample.According to different zymolytes, colour developing is different, judges the kind and the content of (antigen) pathogenic microorganism in the key sample not or antibody.
This method can be prepared by single job, the synchronous detection multiple immune indexes, and detection speed is fast, saves human and material resources, reduces and detects cost, is easy to promote in basic unit.
Claims (4)
1, a kind of method of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) specific antibody with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in patient's the sample adding enzyme linked plate holes;
(3) kind and the quantity of specific antibody in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antibodies of quantity, fully wash plate after the reaction;
(2) preparation control group
(1) method is identical with the step (1) of preparation test sample;
(2) kind and the quantity of specific antibody in the corresponding step (1) join the known antigens of identical kind and quantity in the enzyme linked plate holes,
(3) method is identical with the step (3) of preparation test sample;
(3) control test
A kind of enzyme in corresponding test sample and the control group step (3) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
2, a kind of method of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) specific antibody with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in patient's the sample adding enzyme linked plate holes, and simultaneously
(3) kind and the quantity of specific antibody in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known antigens of quantity, fully wash plate after the reaction;
(2) preparation control group
(1) method is identical with the step (1) of preparation test sample;
(2) method is identical with the step (3) of preparation test sample;
(3) control test
A kind of enzyme in corresponding test sample step (3) and the control group step (2) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
3, a kind of method of enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes comprises the following order step:
Preparation test sample and control group:
(1) preparation test sample:
(1) the antigen pathogen with two or more is coated in the same enzyme linked plate holes, adsorbs back 4 ℃ and dries in the shade;
(2) extract in patient's the sample adding enzyme linked plate holes;
(3) kind and the quantity of antigen pathogen in the corresponding step (1) will join in the enzyme linked plate holes with the not identical kind of enzyme labeling of the same race and the known specific antibody of quantity, fully wash plate after the reaction;
(2) preparation control group
(1) method is identical with the step (1) of preparation test sample;
(2) method is identical with the step (3) of preparation test sample;
(3) control test
A kind of enzyme in corresponding test sample step (3) and the control group step (2) adds a kind of substrate simultaneously earlier, and colour developing is washed plate after relatively detecting, and corresponding afterwards another kind of enzyme adds another kind of substrate, and colour developing is washed plate after relatively detecting, and finishes to detecting successively.
4, according to the method for claim 1,2 or 3 described enzyme-linked immunosorbent ELISA synchronous fast assay multiple immune indexes, it is characterized in that: described elisa plate is a polystyrene board.
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| CN200310114530.0 | 2003-12-20 | ||
| CNA2003101145300A CN1554948A (en) | 2003-12-20 | 2003-12-20 | Method for synchronous quick detecting multiple immune indexes by enzyme linked immunosorbent assay ELISA |
| CN 200410104265 CN1673745A (en) | 2003-12-20 | 2004-12-18 | Process for enzyme-linked immunosorbent ELISA synchronous fast assay method for multiple immune indexes |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102735851A (en) * | 2012-07-13 | 2012-10-17 | 江苏省农业科学院 | Mycoplasma hyopneumoniae multi-recombination antigen ELISA (enzyme-linked immunosorbent assay) detection kit |
| CN102928597A (en) * | 2012-10-30 | 2013-02-13 | 北京维德维康生物技术有限公司 | Enzyme-linked immuno sorbent assay (ELISA) method for detecting sulfonamides and quinolones simultaneously and kit thereof |
| CN103175961A (en) * | 2013-03-12 | 2013-06-26 | 中国农业大学 | Enzyme linked immunosorbent assay kit for detecting 21 sulfonamides and 11 quinolones and application of enzyme linked immunosorbent assay kit |
| CN105675892A (en) * | 2008-04-29 | 2016-06-15 | 赛凯米迪克斯股份有限公司 | Solid phase multi-analyte assay |
| CN107991481A (en) * | 2016-10-27 | 2018-05-04 | 武汉科前生物股份有限公司 | It is a kind of to detect porcine pseudorabies virus and the bigeminy blocking ELISA antibody assay kits of foot and mouth disease virus and its application |
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2004
- 2004-12-18 CN CN 200410104265 patent/CN1673745A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105675892A (en) * | 2008-04-29 | 2016-06-15 | 赛凯米迪克斯股份有限公司 | Solid phase multi-analyte assay |
| US10539580B2 (en) | 2008-04-29 | 2020-01-21 | Psychemedics Corporation | Solid phase multi-analyte assay |
| CN102735851A (en) * | 2012-07-13 | 2012-10-17 | 江苏省农业科学院 | Mycoplasma hyopneumoniae multi-recombination antigen ELISA (enzyme-linked immunosorbent assay) detection kit |
| CN102928597A (en) * | 2012-10-30 | 2013-02-13 | 北京维德维康生物技术有限公司 | Enzyme-linked immuno sorbent assay (ELISA) method for detecting sulfonamides and quinolones simultaneously and kit thereof |
| CN102928597B (en) * | 2012-10-30 | 2015-06-10 | 北京维德维康生物技术有限公司 | Enzyme-linked immuno sorbent assay (ELISA) method for detecting sulfonamides and quinolones simultaneously and kit thereof |
| CN103175961A (en) * | 2013-03-12 | 2013-06-26 | 中国农业大学 | Enzyme linked immunosorbent assay kit for detecting 21 sulfonamides and 11 quinolones and application of enzyme linked immunosorbent assay kit |
| CN107991481A (en) * | 2016-10-27 | 2018-05-04 | 武汉科前生物股份有限公司 | It is a kind of to detect porcine pseudorabies virus and the bigeminy blocking ELISA antibody assay kits of foot and mouth disease virus and its application |
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