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Enzyme linked immunosorbent assay kit for specific quantitative detection of foot and mouth disease O type Guangxi strain antigen and application thereof

An enzyme-linked immunosorbent reagent, a technology for foot-and-mouth disease, used in accurate detection and analysis, enzyme-linked immunosorbent assay kit, specific and rapid field of foot-and-mouth disease O-type Guangxi strain antigen, can solve problems such as difficulties, and achieve the effect of shortening the detection period

Active Publication Date: 2016-03-30
JINYUBAOLING BIO PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it becomes very difficult to prepare monoclonal antibodies by direct immunization with viruses, and to screen them to obtain specific antibodies against specific serotypes, especially different strains of the same serotype

Method used

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  • Enzyme linked immunosorbent assay kit for specific quantitative detection of foot and mouth disease O type Guangxi strain antigen and application thereof
  • Enzyme linked immunosorbent assay kit for specific quantitative detection of foot and mouth disease O type Guangxi strain antigen and application thereof
  • Enzyme linked immunosorbent assay kit for specific quantitative detection of foot and mouth disease O type Guangxi strain antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064]Embodiment 1, prepare the enzyme-linked immunosorbent assay kit of foot-and-mouth disease O type Guangxi strain (O / GX / 09-7 strain)

[0065] One, prepare the specific monoclonal antibody of foot-and-mouth disease O type Guangxi strain (O / GX / 09-7 strain)

[0066] Prepare the specific monoclonal antibody of foot-and-mouth disease O type Guangxi strain (O / GX / 09-7 strain), and concrete method comprises the following steps:

[0067] 1. Preparation of FMD O-type Guangxi strain (O / GX / 09-7 strain) specific antigen

[0068] Get purified and concentrated foot-and-mouth disease O-type Guangxi strain (O / GX / 09-7 strain) purified antigen (commercial product provided by Jinyu Baoling Biopharmaceutical Co., Ltd.), the concentration is greater than 100 μg / mL, and carry out 2-DE electrophoresis, the method is as follows :

[0069] The first isoelectric focusing, specific operation steps: 1) Take the hydration loading buffer (Urea480mg / mL, CHAPS20mg / mL, IPGBuffer5μl / mL, DTT2.8mg / mL, trace...

Embodiment 2

[0139] The using method of the ELISA quantitative assay kit of embodiment 2, foot-and-mouth disease O type Guangxi strain (O / GX / 09-7 strain)

[0140] Detecting the sample with the kit of the present invention comprises the following steps:

[0141] 1) Use the purified and inactivated solution of 146S of the Guangxi strain of foot-and-mouth disease O type (O / GX / 09-7 strain) and the sample diluent provided by the kit to prepare standard products. The concentrations of the standard products are: 0, 2, 4, 8, 16 , 32, 64 ng / mL. The sample to be tested is also diluted 4-8 gradients with the sample diluent.

[0142] 2) Take out the required slats, put the remaining slats into a ziplock bag, and store them at 2-8°C.

[0143] 3) Add 100 μl of serial dilution points of standard product, positive quality control, and sample serial dilution points to the wells respectively. Gently vibrate for 10-20 seconds to mix well, and paste on the board.

[0144] 4) Place in a 37°C incubator and ...

Embodiment 3

[0182] Embodiment 3, use the enzyme-linked immunosorbent assay kit quantitative assay reagent detection sample of foot-and-mouth disease O type Guangxi strain (O / GX / 09-7 strain) of the present invention

[0183] Detect the O / GX / 09-7 strain 146S antigen concentration in samples and finished vaccines of the different production stages of the foot-and-mouth disease O-type Guangxi strain (O / GX / 09-7 strain) with the kit of embodiment 1 and the using method of embodiment 2 (for quality control purposes). These samples are all from Jinyu Baoling Biological Pharmaceutical Co., Ltd. Among them, the three finished vaccines with serial numbers 5, 6, and 7 were tested after demulsification according to the following method: add 20 mL of n-amyl alcohol to 100 mL of vaccine, stir for 1 minute and then rest for 30 minutes to separate the upper and lower layers, and take the water phase for detection.

[0184] The results are shown in Table 7. It can be seen that the kit can not only detect ...

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Abstract

The invention discloses an enzyme linked immunosorbent assay kit for specific quantitative detection of a foot and mouth disease O type Guangxi strain (O / GX / 09-7 strain) antigen and an application thereof. The kit is a double-antibody sandwich-method enzyme-linked immune quantitative detection kit prepared from an O / GX / 09-7 strain specific monoclonal antibody, and the content of the O / GX / 09-7 strain antigen in a sample is quantitatively detected through detection of changes of signals generated by an enzyme catalyzed substrate. The detection using the kit does not generate cross interference with other current foot and mouth disease vaccine strains such as O / Mya98 / XJ / 2010 strains, O / Mya98 / BY / 2010, O / JMS / 2000, OZK / 93 strains, Asia1 / JSL / 06 and type A A / WH / 09, and the method can simply, rapidly, sensitively and stably detect the content of the foot and mouth disease O type Guangxi strain antigen and specifically distinguish a foot and mouth disease O type Guangxi strain and other strains.

Description

technical field [0001] The invention belongs to the technical field of enzyme-linked immunoimmunoassay detection and analysis, in particular to an enzyme-linked immunosorbent assay kit for specific and quantitative determination of the antigen of foot-and-mouth disease O-type Guangxi strain (O / GX / 09-7 strain), which is suitable for foot-and-mouth disease O-type Guangxi strain ( O / GX / 09-7 strain) antigen specific, rapid and accurate detection analysis. Background technique [0002] Foot and Mouth Disease Virus (Foot and Mouth Disease Virus, FMDV) belongs to Picornaviridae, Foot and Mouth Disease Virus genus, the virus has 7 serotypes, no cross-protective reaction among the various types. At present, the main popular ones in China are O type, Asian type 1 (such as JSL / 06 strain) and A type (A / WH / 09 strain). Among them, type O includes multiple strains, namely Xinjiang strain (O / Mya98 / XJ / 2010 strain), Jiamusi strain (O / JMS / 2000 strain), Myanmar 98 strain (O / Mya98 / BY / 2010 strai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531G01N33/543G01N21/31
Inventor 刘国英郑金来吴园园任旭荣范秀丽苍枫刘飞
Owner JINYUBAOLING BIO PHARMA CO LTD
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