Detection method and detection kit for porcine type A foot-and-mouth disease virus specific IgA antibody

A technology of foot-and-mouth disease virus and detection kit, which is applied to measuring devices, instruments, scientific instruments, etc., to achieve the effects of low manpower consumption, easy operation, and low stress

Inactive Publication Date: 2017-05-24
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no kit that can effectively detect FMD virus-specific IgA antibody on the market. In view of the in-depth research on mucosal immune mechanism and mucosal immune vaccine in recent years, an ELISA for detecting the level of FMD virus-specific IgA antibody in mucosal secretions was developed. The kit can more accurately reflect the FMD virus infection and mucosal immune status, and at the same time use this method to monitor the dynamic changes of FMD virus-specific IgA antibodies, providing a theoretical basis for the evaluation of the FMD mucosal immune effect and the optimization of the immune program

Method used

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  • Detection method and detection kit for porcine type A foot-and-mouth disease virus specific IgA antibody
  • Detection method and detection kit for porcine type A foot-and-mouth disease virus specific IgA antibody
  • Detection method and detection kit for porcine type A foot-and-mouth disease virus specific IgA antibody

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Embodiment 1

[0038] Preparation method of the kit:

[0039]1. Preparation of coating antigen and selection of optimal coating concentration:

[0040] The 636bP fragment of the A-type FMDV-VP1 protein was cloned into the pET-30a (+) prokaryotic expression vector. After digestion and sequencing, it was transformed into E.coli BL-21 (E3), and the VP1 protein was induced by IPTG. Proteins were purified using the Ni-Agarose His-Tag Protein Purification Kit. The purified product expressed by the prokaryotic expression system was serially diluted with carbonate buffer 7μg / mL, 3.5μg / mL, 1.75μg / mL, 0.88μg / mL, 0.44μg / mL, 0.22μg / mL, each Two columns were coated with each concentration, 100 μL / well was added to a 96-well microtiter plate, and coated overnight at 4°C. The results showed that when the coating antigen concentration was 3.5μg / mL, the positive value OD450 value was greater than 1.0, and the value of P / N was the largest, so this concentration was determined to be the optimal coating conce...

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Abstract

The invention discloses a detection method for a porcine type A foot-and-mouth disease virus specific IgA antibody; a prokaryotic expression system-expressed foot-and-mouth disease virus VP1 protein is used as a coating antigen, a mouse anti porcine IgA monoclonal antibody is used as a secondary antibody, an enzyme-labeled goat anti mouse IgG antibody is used as an indicator, a to-be-tested sample is subjected to reactions with the coating antigen, the monoclonal antibody and the enzyme-labeled antibody successively, and then is compared with a reference positive sample and a negative sample, and the result can be used for evaluating the level of the porcine foot-and-mouth disease virus specific IgA antibody; and a detection kit is provided. The detection method has the beneficial effects that the invention provides an effective method for evaluating the immune effect of foot-and-mouth disease mucosa and provides a new method for early diagnosis of infection of foot-and-mouth disease, and the foot-and-mouth disease virus specific IgA antibody in porcine nasal swabs can be quickly and accurately detected, sample collection operation is simple, manpower consumption is low, and the stress on animals is small.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for detecting porcine type A foot-and-mouth disease virus-specific IgA antibody and a detection kit thereof. Background technique [0002] Foot-and-Mouth Disease (FMD) is an acute, febrile, highly contactable and fast-moving animal disease caused by the Foot-and-Mouth Disease Virus of the family Picornaviridae. Decreased performance and high mortality in sick litters. Infection targets are mainly pigs, cattle, sheep and other livestock species and other cloven-hoofed animals, and there are more than 70 susceptible animals. Once the disease breaks out, it will bring great economic losses to the breeding industry and international animal product trade. The International Organization for Animal Health (OIE) lists it at the top of the list of 15 Class A animal diseases, and it must be reported once it occurs in my country. One of a class of animal infectious di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56983G01N33/577
Inventor 潘丽王园园吕建亮张中旺郑华斌王永录张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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