Pig porcine deltacoronavirus N protein indirect ELISA antibody detection method and kit thereof

A coronavirus and detection method technology, which is applied in the field of porcine D-coronavirus N protein indirect ELISA antibody detection method and its kit, can solve the problems of less research on PDCoV antibody detection methods, and achieve low cost, high specificity and Sensitive, time-consuming effect

Active Publication Date: 2019-04-19
GUANGXI VETERINARY RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the diagnostic method for PDCoV is mainly RT-PCR method, and there are few studies on the detection method for PDCoV antibody

Method used

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  • Pig porcine deltacoronavirus N protein indirect ELISA antibody detection method and kit thereof
  • Pig porcine deltacoronavirus N protein indirect ELISA antibody detection method and kit thereof
  • Pig porcine deltacoronavirus N protein indirect ELISA antibody detection method and kit thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0024] Establishment of an indirect ELISA method for detection of porcine D-coronavirus N protein antibody

[0025] 1. Materials, Reagents and Instruments

[0026] 1.1 Main reagents

[0027] PBS powder: product of Boster;

[0028] Skimmed milk powder: product of solarbio company;

[0029] HRP-labeled goat anti-pig IgG antibody: product of Earthox Company in the United States;

[0030] One-component TMB substrate chromogenic solution: product of Tiangen Company;

[0031] 96-well ELISA plate: Constar company product;

[0032] Recombinant N protein: self-made (encoded by the gene base sequence of SEQ.ID.No.1 in the sequence table, and has the amino acid sequence of SEQ.ID.No.2).

[0033] 1.2 Main instruments

[0034] Water-proof electric heating constant temperature incubator (Shanghai Yuejin), ultraviolet spectrophotometer (Tianpu), microplate reader (Bole), etc.

[0035] 1.3 Preparation of related reagents for indirect ELISA

[0036] 1) Coating solution: use 0.05M PH9.6...

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Abstract

The invention discloses a pig porcine deltacoronavirus N protein indirect ELISA antibody detection kit. The pig porcine deltacoronavirus N protein indirect ELISA antibody detection kit comprises a coated ELISA plate, and the coated ELISA plate takes recombinant N protein as a coated antigen. According to the detection kit, a corresponding ELISA detection method is further built and a diagnosis tool with practical value is provided for PECoV detection. By the adoption of the pig porcine deltacoronavirus N protein indirect ELISA antibody kit, the method for identifying the antibody generated byinfection of pig porcine deltacoronavirus is built, has high specificity and sensitivity, and is easy to operate, short in consumed time and low in cost; detection can be conducted in large batch, andthe detection kit and method have great significance on prevention and purification of pig porcine deltacoronavirus.

Description

technical field [0001] The invention belongs to the technical field of porcine D-coronavirus, in particular to a method for detecting porcine D-coronavirus N protein indirect ELISA antibody and a kit thereof. Background technique [0002] Porcine deltacoronavirus (porcine deltacoronavirus, PDCoV) was first discovered in 2012. Woo et al. from the University of Hong Kong detected porcine deltacoronavirus from dead mammals. Coronaviruses are divided into four subgroups, A, B, C, and D, and the PDCoV that infects pigs is the only PDCoV that infects non-birds. Since then, the prevalence of PDCoV has been reported successively in the United States, China, South Korea and many other countries. Similar to PEDV and TGEV, PDCoV mainly causes intestinal diseases in pigs, leading to diarrhea, vomiting and dehydration in infected pigs, and the morbidity and mortality can be as high as 50%-100%, especially in suckling piglets, which seriously threatens pigs. The health of the flock has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/535
CPCG01N33/535G01N33/56983G01N33/6803
Inventor 刘磊卢冰霞秦毅斌陈忠伟赵武何颖李斌梁家幸周英宁段群棚蒋冬福卢敬专
Owner GUANGXI VETERINARY RES INST
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