猪流行性腹泻病毒与猪伪狂犬病病毒二重RT-PCR检测方法的建立及应用

doi:10.16431/j.cnki.1671-7236.2016.09.034

Abstract

Abstract:

In order to establish an assay for detecting porcine epidemic diarrhea virus (PEDV) and pseudorabies virus (PRV),two pairs of primers were designed basing on the M gene of PEDV and gE gene of PRV,respectively.The total RNA of standard PEDV and PRV strains were used as templates to establish the duplex RT-PCR assay.The specificity,sensitivity,repetition and clinic detection of the established assay were tested.The result revealed that the threshold of duplex RT-PCR was 10 TCID₅₀/mL of PEDV and PRV,and no products were amplified from the cell or the nucleic acid of other 7 kinds of pathogenic viral or bacterial microorganism.The detection results for 26 clinical suspicious PEDV or PRV infected pigs were consistent with the results tested by sequencing.This study suggested that the duplex RT-PCR method was highly specific,repeatable and sensitive,and was suitable for clinic rapid differential diagnosis of PEDV and PRV.

Key words: porcine epidemic diarrhea virus; pseudorabies virus; duplex RT-PCR

CLC Number:

S858.28

SUN Yan-qin, MA Zhen-yuan, YAN Ruo-qian, WANG Dong-fang, CAO Wei-wei, GUO Yu-pei. Establishment and Application of Duplex RT-PCR Assay for Detection of Porcine Epidemic Diarrhea Virus and Pseudorabies Virus[J]. , 2016, 43(9): 2455-2460.

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Establishment and Application of Duplex RT-PCR Assay for Detection of Porcine Epidemic Diarrhea Virus and Pseudorabies Virus

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