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Porcine D-coronavirus N protein indirect ELISA antibody detection method and its kit

A coronavirus and antibody detection technology, applied in biological tests, measurement devices, material inspection products, etc., can solve the problems of less research on PDCoV antibody detection methods, and achieve low cost, short time consumption, high specificity and sensitivity Effect

Active Publication Date: 2022-06-03
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Porcine D-coronavirus N protein indirect ELISA antibody detection method and its kit
  • Porcine D-coronavirus N protein indirect ELISA antibody detection method and its kit
  • Porcine D-coronavirus N protein indirect ELISA antibody detection method and its kit

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

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

[0025] 1. Materials, reagents and instruments

[0026] 1.1 Main reagents

[0027] PBS powder: Boster product;

[0028] Skimmed milk powder: solarbio product;

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

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

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

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

[0033] 1.2 Main instruments

[0034] Waterproof electric heating constant temperature incubator (Shanghai Yuejin), UV spectrophotometer (Tempo), microplate reader (Bole), etc. 1.3 Preparation of indirect ELISA related reagents

[0035] 1) Coating solution: use 0.05M PH9.6 Na 2 CO 3 -NaHCO 3 Buffer, weigh...

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Abstract

The invention discloses an indirect ELISA antibody detection kit for porcine D-coronavirus N protein, which comprises a coated microplate plate, and the coated microplate uses recombinant N protein as a coated antigen. Accordingly, the corresponding ELISA detection method is also established, which provides a valuable diagnostic tool for the detection of PDCoV. Application of the present invention can establish the method for identifying the antibody produced by porcine D-coronavirus infection, which has high specificity and sensitivity. Control and purification are of great significance.

Description

technical field [0001] The invention belongs to the technical field of porcine deltacoronavirus, in particular to a porcine deltacoronavirus N protein indirect ELISA antibody detection method and a kit thereof. Background technique [0002] Porcine deltacoronavirus (PDCoV) was first discovered in 2012. Woo et al. of the University of Hong Kong detected porcine deltacoronavirus from dead mammals. Coronaviruses are divided into four subgroups, A, B, C, and D, and the pig-infecting delta-coronavirus (PDCoV) is the only delta-coronavirus that infects non-avian species. Since then, the prevalence of PDCoV has been successively reported in many countries such as the United States, China, and South Korea. 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 rate can be as high as 50%-100%, especially in suckling piglets, which is a serious threat to pigs. The heal...

Claims

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Application Information

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Patent Type & Authority Patents(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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