Primers for differential diagnosis of porcine epidemic diarrhea virus virulent strain and attenuated strain and detection kit thereof

A technology for differential diagnosis of porcine epidemic diarrhea virus, applied in the field of primers and detection kits for differential diagnosis of strong and weak strains of porcine epidemic diarrhea virus, can solve the problems of time-consuming, high cost, and long time required for gene sequencing technology , to achieve high sensitivity and good effect

Active Publication Date: 2015-04-08
JIANGSU ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, other RT-PCR methods cannot solve this problem; gene sequencing technology takes a long time, and is time-consuming, laborious and costly

Method used

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  • Primers for differential diagnosis of porcine epidemic diarrhea virus virulent strain and attenuated strain and detection kit thereof
  • Primers for differential diagnosis of porcine epidemic diarrhea virus virulent strain and attenuated strain and detection kit thereof

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Experimental program
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Effect test

Embodiment 1

[0055] 1. Design and synthesis of primers

[0056] According to the nucleotide sequence of porcine epidemic diarrhea virus M gene, primers were designed using Primer 5.0, and the primer sequences of the following sequences were obtained through a large number of screenings:

[0057] The sequence of the M gene forward primer (PEDV-MF) is SEQ ID NO.1, and the sequence of the M gene reverse primer (PEDV-MR) is SEQ ID NO.2. The composition of the specific sequence is as follows:

[0058] SEQ ID NO.1: 5'-TATTCCCGTTGATGAGGTGATTG-3'

[0059] SEQ ID NO.2: 5'-GCTGAATAGTCGCCGTGTTTTG-3'

[0060] According to the characteristic that the ORF3 gene of the attenuated porcine epidemic diarrhea virus strain lacks 49nt or 51nt, primers were designed with Primer 5.0 for the ORF3 gene and its adjacent gene sequences, and the primer sequences of the following sequences were obtained after extensive screening:

[0061] The sequence of the ORF3 gene forward primer (PEDV-ORF3F) is SEQ ID NO.3, an...

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Abstract

The invention relates to primers for differential diagnosis of a porcine epidemic diarrhea virus virulent strain and attenuated strain and a detection kit thereof, and belongs to the field of biotechnology. Total RNA is used as a template, the detection primers for differential diagnosis of the porcine epidemic diarrhea virus virulent strain and attenuated strain are used for RT-PCR amplification, and after the reaction is finished, a result is judged according to the amplified fragment number and the amplified fragment molecular weight range. The primers have good specificity, the detection method is fast and simple, the accuracy is high, and a guarantee is provided for the differential diagnosis of the porcine epidemic diarrhea virus virulent strain and attenuated strain.

Description

technical field [0001] The invention relates to primers for differential diagnosis of strong and weak strains of porcine epidemic diarrhea virus and a detection kit thereof, belonging to the field of biotechnology. Background technique [0002] Porcine epidemic diarrhea virus (PEDV) is a member of the Coronaviridae family. Clinically, PEDV can cause porcine epidemic diarrhea (porcine epidemic diarrhea, PED), which is characterized by diarrhea, vomiting, dehydration and high lethality in suckling piglets. PED was first reported in Belgium and the UK in 1978 (Pensaert M B, de Bouck P. A new coronavirus—like particle associated with diarrhea in swine. Arch Virol, 1978, 58: 243-247). Its genome is a single-molecule linear positive-sense single-stranded RNA with a size of about 28kb. Its 5' end is capped and its 3' end is a polyA tail, which is infectious. The genome contains 5'-UTR, 3'-UTR and at least 7 ORFs. Four of these seven ORFs encode structural proteins: spike protein...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143
Inventor 倪艳秀何孔旺郭容利李彬俞正玉茅爱华吕立新祝昊丹周俊明温立斌张雪寒王小敏汪伟胡屹屹
Owner JIANGSU ACAD OF AGRI SCI
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