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Multiplex RT-PCR detection primer for porcine delta coronavirus, porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus

A porcine epidemic diarrhea and RT-PCR technology, applied in the field of virus detection, can solve the problems of complicated operation and difficulty in effectively distinguishing virus infection, and achieve the effect of rapid differential diagnosis

Active Publication Date: 2016-04-13
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, these methods are more complicated to operate, and it is difficult to effectively distinguish the specific virus infection

Method used

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  • Multiplex RT-PCR detection primer for porcine delta coronavirus, porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus
  • Multiplex RT-PCR detection primer for porcine delta coronavirus, porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus
  • Multiplex RT-PCR detection primer for porcine delta coronavirus, porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus

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Embodiment Construction

[0028] According to the N gene sequence of HKU15-155 strain of PDCoV, the N gene sequence of H155 strain of TGEV, and the M gene sequence of JXNC1302 strain of PEDV registered in GenBank, the present invention obtains its highly conserved specific sequence through sequence analysis, and according to the specificity conservation Two pairs of specific primers were designed for the sequence. On this basis, a multiplex RT-PCR detection method for PDCoV, TGEV and PEDV was established, and the method was evaluated and analyzed. The samples of disease materials submitted for inspection in Henan area were detected and analyzed, and the reliability of the method was further verified in practice, so as to provide a theoretical basis for veterinary clinical diagnosis and treatment.

[0029] Primers for multiplex RT-PCR detection of porcine delta coronavirus, porcine epidemic diarrhea virus, and porcine transmissible gastroenteritis virus:

[0030] The primer sequences of porcine deltaco...

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Abstract

The invention discloses a multiplex RT-PCR detection primer for a porcine delta coronavirus, a porcine epidemic diarrhea virus and a porcine transmissible gastroenteritis virus. The minimum detection capacity of the multiplex RT-PCR for the three viruses is 4.05*10<1> copies / microliter, 4.52*10<3> copies / microliter and 5.47*10<3> copies / microliter respectively. The amplification results for a porcine parvovirus (PPV) and a porcine pseudorabies virus (PRV) are both negative. The multiplex RT-PCR detection results of 57 clinical samples show that one sample is infected with the three viruses at the same time, 11 samples are infected with the PDCoV, 15 samples are infected with the PEDV, one sample is infected with the TGEV, five samples are infected with the PDCoV and the PEDV, and one sample is infected with the PDCoV and the TGEV.

Description

technical field [0001] The invention relates to a virus detection method, in particular to multiple RT-PCR detection primers and detection methods for porcine delta coronavirus, porcine epidemic diarrhea virus and porcine transmissible gastroenteritis virus. Background technique [0002] Porcine Deltacoronavirus (Porcine Deltacoronavirus, PDCoV) belongs to Nidovirales (Nidovirales), Coronaviridae (Coronaviridae), Coronavirus (Coronavirus) Deltacoronavirus (Coronavirus), is a single-stranded positive-sense RNA virus with an envelope. As early as 2012, there were reports of PDCoV in the articles published by Woo et al.: They detected Delta coronavirus in mammalian and poultry disease materials collected in Guangdong and Hong Kong, among which it was detected in pig feces samples (169 samples) Nearly 10% of PDCoVs. In early 2014, a new porcine enteric coronavirus PDCoV was detected and reported for the first time in Ohio, USA. By the end of 2014, PDCoV infection cases had been...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/09
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143C12Q2521/107
Inventor 胡慧曹贝贝张君涛郑兰兰魏战勇梁秀丽周雍
Owner HENAN AGRICULTURAL UNIVERSITY
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