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ELISA detection method and kit for PEDV specific SIgA antibody in sow colostrum

A technology of detection kits and detection methods, which can be applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as loss, high mortality, and economic troubles in pig farming.

Active Publication Date: 2020-05-19
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Porcine epidemic diarrhea (Porcine epidemic diarrhea, PED) is a disease that occurs in the intestinal tract of pigs and can cause pigs to die after watery diarrhea, vomiting, and dehydration. The disease has the characteristics of acute onset, rapid spread, and high mortality. Bring huge troubles to the pig industry and cause serious economic losses

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  • ELISA detection method and kit for PEDV specific SIgA antibody in sow colostrum
  • ELISA detection method and kit for PEDV specific SIgA antibody in sow colostrum
  • ELISA detection method and kit for PEDV specific SIgA antibody in sow colostrum

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Embodiment Construction

[0037] 1. ELISA detection method for PEDV-specific SIgA antibody in sow colostrum

[0038] 1.1 Antigen coating: Add the antigen PEDV virus particles and coating solution to the microtiter plate for coating. The PEDV virus particles coated in each well are about 10 5, coated at 4°C for at least 12 hours; then washed with PBST to remove unbound antigens and impurities;

[0039] 1.2 Blocking: Add PBS containing 1% BSA by volume as a blocking solution, block at 37°C for 2 hours, then discard the blocking solution, and wash away the residual blocking solution with PBST;

[0040] 1.3 Add antibody: first add 10 times diluted colostrum, incubate at 37°C for 1 hour, wash with PBST after reaction; then add 1000 times diluted SC rabbit polyclonal antibody, incubate at 37°C for 1 hour, wash with PBST after reaction; then add 2000 times Diluted HRP-labeled goat anti-rabbit IgG, incubated at 37°C for 1 hour, washed with PBST after reaction;

[0041] 1.4 Color development and measurement: ...

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Abstract

The invention discloses an ELISA detection method for a PEDV specific SIgA antibody in sow colostrum. According to the method, technologies of gene cloning, prokaryotic expression, polyclonal antibodypreparation and the like are applied; the method comprises the following steps: cloning from the sow colostrum to obtain a specific component SC fragment of SIgA and constructing a prokaryotic expression vector to express the fragment; then coating the prepared rabbit anti-pig SC polyclonal antibody with PEDV virus, sealing, sequentially adding a primary antibody, a secondary antibody and a tertiary antibody and incubating for 1h; and finally adding a stop solution to measure the OD450nm value. Accordingly, the invention also provides a corresponding kit. The method has the characteristics ofeconomy, convenience, quickness and accuracy, provides a reliable method for further exploring the dynamic change rule of the PEDV specific SIgA antibody in the sow milk, evaluating the PEDV mucosalimmune effect and optimizing the immune procedure and also provides a scientific basis for preventing infection of the PEDV to suckling piglets.

Description

technical field [0001] The invention belongs to the technical field of animal antibody detection, in particular to an ELISA detection method for PEDV-specific SIgA antibody in sow colostrum and a kit thereof. Background technique [0002] Porcine epidemic diarrhea (Porcine epidemic diarrhea, PED) is a disease that occurs in the intestinal tract of pigs and can cause pigs to die after watery diarrhea, vomiting, and dehydration. The disease has the characteristics of acute onset, rapid spread, and high mortality. Bring huge puzzlement to pig industry and cause serious economic loss. PEDV mainly infects suckling piglets, and its infection and incidence are closely related to the age of suckling piglets, especially young piglets (below 10 days old) are most susceptible to infection, and their morbidity and mortality can reach 100%. Therefore, improving the resistance of suckling piglets to PEDV is very important to prevent PED. Mucosal immunity, as the first line of defense ag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/165
Inventor 欧阳康王若木杜琛陆颖黄伟坚韦祖樟陈樱
Owner GUANGXI UNIV