Primer set for distinguishing porcine epidemic diarrhea virus typical and variant strains and detection kit

A porcine epidemic diarrhea and detection kit technology, which is applied in the determination/testing of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of cumbersome detection methods, low sensitivity, expensive instruments, etc., and achieve cost Low, Guaranteed Accuracy, High Accuracy Effect

Inactive Publication Date: 2019-05-10
JILIN ZHENGYE BIOLOGICAL PROD
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Problems solved by technology

However, the current detection method not only has many disadvantages such as cumbersome operation, long time, low sensitivity and expensive equipment, but also brings great difficulties to the accurate diagnosis of PEDV in the face of the current complex epidemic situation where classic and mutant strains of PEDV are co-populated; therefore, There is an urgent need to develop a method with good stability and high sensitivity that can simultaneously identify PEDV classic and variant strains

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  • Primer set for distinguishing porcine epidemic diarrhea virus typical and variant strains and detection kit
  • Primer set for distinguishing porcine epidemic diarrhea virus typical and variant strains and detection kit
  • Primer set for distinguishing porcine epidemic diarrhea virus typical and variant strains and detection kit

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Embodiment 1

[0032] Example 1 Establishment of Differential Diagnosis Method for Porcine Epidemic Diarrhea Virus Classical and Variant Strains

[0033] 1 Test method

[0034] 1.1. Primer design

[0035] The present invention is based on a large number of sequence comparisons of strains such as the classic strain representative CV777 (accession number AF353511) and the variant strain representative AH2012 (accession number KC210145) published by GenBank in NCBI, and based on the 175 bp of the S gene of the two types of strains Stable differential characteristics, designed multiple sets of Nano-nest PCR detection primer sets, the specific sequences are as follows:

[0036] The first set of Nano-nest PCR detection primer set:

[0037] Nest-F1: 5'-TTTAGGCGGTTTCTTTTCA-3' (SEQ ID No.1);

[0038] Nest-R1: 5'-TTACAAACRCCATCSATC-3' (SEQ ID No.2);

[0039] Nest-F2: 5'-CAGTTTCCHAGCATYAAA-3' (SEQ ID No.3);

[0040] Nest-R2: 5'-TACCATCCTCACCAGCAC-3' (SEQ ID No. 4).

[0041] The second set of Nano...

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Abstract

The invention discloses a Nano-nest PCR detection kit capable of distinguishing porcine epidemic diarrhea virus typical and variant strains. Firstly, two pairs of specific primers are obtained throughsieving, only amplify porcine epidemic diarrhea viruses, has no specific amplification for porcine transmissible gastroenteritis viruses, rotaviruses, hog cholera viruses, pseudorabies viruses and porcine reproductive and respiratory syndrome viruses, and can guarantee the accuracy of a detection result; on the basis, the invention establishes a Nano-nest PCR detection method capable of distinguishing porcine epidemic diarrhea virus typical and variant strains. The method is high in accuracy and specificity and low in cost, the sensitivity is 100 times that of common PCR, and the method is suitable for epidemiological survey of porcine epidemic diarrhea viruses and also suitable for PEDV accurate treatment and prevention and control.

Description

technical field [0001] The present invention relates to the identification of porcine epidemic diarrhea virus classic and variant strains, in particular to a primer set and Nano-nest PCR detection kit for identifying porcine epidemic diarrhea virus classic and variant strains, belonging to porcine epidemic diarrhea virus classic and variant strains The field of molecular identification of virus strains. Background technique [0002] Porcine epidemic diarrhea virus (Porcine epidemic diarrhea virus, PEDV) is a member of the Coronaviridae family and the genus Alphacoronavirus, which can cause clinical symptoms such as acute diarrhea, vomiting and dehydration in piglets, and eventually lead to death of sick pigs due to electrolyte balance disorders. Gastrointestinal viruses have caused major economic losses to the world's pig industry. PEDV is a single-stranded positive-sense RNA virus with an envelope and a crown shape. Its genome is 28 kb in length and encodes three nonstruct...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6848C12N15/11
Inventor 朱俊辉陈立思何玉友王国辉廉维杨泽彬王石李长艳白杨贾小营
Owner JILIN ZHENGYE BIOLOGICAL PROD
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