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Protein chip for diagnosing porcine virus diarrhea

A porcine viral diarrhea and protein chip technology is applied in the field of bioinformatics, which can solve problems such as economic losses

Inactive Publication Date: 2015-03-25
周丽华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PED first occurred in Europe, it has become a serious infectious disease in the pig industry in Asian countries including China, South Korea, Japan, the Philippines and Thailand, causing huge economic losses

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0012] 1. Selection of polypeptide position and length

[0013] Using bioinformatics methods to analyze the protein sequences with characteristic epitopes in the extracellular parts of the three viral membrane proteins (M protein and S protein), and then analyze the position and homology of these sequences on the protein and membrane, according to According to the corresponding characteristics, a certain length is reasonably selected and sent to Shanghai Bioengineering Co., Ltd. for synthesis.

[0014] 2. Immunization of hens

[0015] The synthesized polypeptide was coupled with BSA, fully emulsified with Freund's complete adjuvant for the first time, multi-point subcutaneous injection to immunize hens, and incomplete adjuvant was used for the second and subsequent immunizations, with a time interval of half a month , After the fourth immunization is completed, the antibody is extracted, the titer is tested, and the chickens whose titer, sensitivity and specificity do not mee...

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Abstract

The invention relates to a protein chip for diagnosing porcine virus diarrhea. Hens are immunized by synthesizing polypeptide aiming at extracellular epitopes of three virus membrane proteins; a pair of IgY antibodies is screened from each virus, so as to prepare a solid-phase support object; the IgY antibodies are combined on the solid-phase support object after one IgY antibody in the pair of antibodies of each virus is incubated; when the solid-phase support object combined with the IgY reacts with a to-be-detected sample, most of virus particles in the sample are specifically combined; the other IgY antibody in the pair of antibodies is added; a secondary antibody marked by alkaline phosphatase is added; a substrate AMPPD is added after being washed and separated; a mesosome AMPD is generated by the AMPPD; the AMPD can get back to a low-energy stable state from a high-energy excited state; meanwhile, photons are emitted; the luminescent intensity is recorded through a light quantum recording system; and the quantity of the virus particles in the to-be-detected sample are calculated from a standard curve.

Description

technical field [0001] The present invention relates to the field of immunodiagnosis, specifically a method of bioinformatics to predict the epitopes of the extracellular parts of three viral membrane proteins that cause viral diarrhea in piglets. According to the specific characteristics of epitopes, structures, and homology, Synthesize a polypeptide of a certain length, immunize hens, and obtain three pairs of IgY antibodies with high sensitivity and specificity, and then prepare a solid phase support, and use the method of making a gene chip to determine the amount of specific proteins on the surface of the three virus particles The number and type of virus particles, and then a protein chip for rapid diagnosis of porcine viral diarrhea. Background technique [0002] Porcine epidemic diarrhea (porcine epidemic diarrhea, PED) is a porcine intestinal infectious disease caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV). At present, it co-infe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/56983G01N33/6842G01N2333/14
Inventor 周丽华
Owner 周丽华
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