Porcine sapovirus (SaV) total immunoglobulin ELISA (Enzyme-Linked Immunosorbent Assay) detection kit and preparation method thereof
A detection kit and technology for Sapporo virus, applied in the field of biomedicine, can solve the problems of no commercial kits for detecting porcine SaY antibody and immature detection methods for swine-derived Sapporo virus, and achieve low reagent cost, high detection sensitivity, The effect of simple operation steps
Active Publication Date: 2016-04-13
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology
[0004] At present, domestic and foreign detection methods for porcine-origin Sapporo virus are immature, and there is no commercial kit for detection of porcine SaY antibody, but mainly focus on the detection of virus morphology and viral RNA.
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[0053] The present invention will be further described below in conjunction with the accompanying drawings. It should be noted that this embodiment is based on the technical solution, and provides detailed implementation and specific operation process, but the protection scope of the present invention is not limited to the present invention. Example.
[0054] An ELISA detection kit for total antibody to porcine Sapporo virus, comprising an ELISA plate containing a SaV coating antigen and a SaV labeled antigen-HRP (horseradish peroxidase) marker, wherein the SaV coating antigen is SaVswCH430 strain P Protein fragment, the SaV marker antigen is the S protein fragment of the SaVswCH430 strain.
[0055] Such as figure 1 Shown, a kind of method for preparing above-mentioned porcine sapovirus total antibody ELISA detection kit comprises the steps:
[0056] S1 preparation of SaV coating antigen and preparation of SaV labeled antigen;
[0057] 1.1) First, PCR amplifies two fragmen...
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Abstract
The invention discloses a porcine sapovirus (SaV) total immunoglobulin ELISA (Enzyme-Linked Immunosorbent Assay) detection kit and a preparation method thereof. The detection kit comprises an Elisa plate containing a SaV coating antigen and a SaV labelled antigen-HRP (Horse Radish Peroxidase) marker. The preparation method comprises the following steps: preparing the SaV coating antigen and preparing the SaV labelled antigen; preparing the SaV labelled antigen-HRP marker; diluting the SaV coating antigen with a carbonate buffer solution according to a certain proportion, adding the diluted SaV coating antigen into the ELISA plate by 100 mumL per hole, and packaging in an aluminum film bag added with a drying agent, and finishing coating. The detection kit is based on a double-sandwich ELISA principle, is simple and convenient in operation steps, is low in reagent cost, is high in detection sensitivity, and is particularly suitable for rapid batch detection of porcine serum specimens. A basis is laid for the investigation of porcine SaV epidemiology.
Description
technical field [0001] The invention relates to the technical field of biomedicine, in particular to an ELISA detection kit for porcine Saapporo virus total antibody and a preparation method thereof. Background technique [0002] Sapovirus (Sapovirus, SaY) belongs to Caliciviridae (Caliciviridae), Sapporo-like virus (Sappora-likevirus), is a single-stranded positive-sense RNA virus. It can cause acute viral gastroenteritis and diarrhea in humans and animals, and is usually transmitted by fecal-oral route. [0003] Pig SaY is widely distributed and has been detected in Europe, Asia and the Americas. It can infect pigs of all ages. Weaned piglets around 1 month old are most susceptible. Adult pigs rarely show clinical symptoms after infection, causing serious damage to the pig industry. certain threats and losses. The latest research shows that there are similarities in genetics and antigenic characteristics between human and animal SaY of different genotypes, and SaY recomb...
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IPC IPC(8): G01N33/569G01N33/535C07K14/08
CPCC07K14/005C12N2770/16022G01N33/535G01N33/56983G01N2333/08
Inventor 兰喜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI