新疆地区4种猪病多重PCR检测方法的建立与初步应用

doi:10.16431/j.cnki.1671-7236.2015.10.015

Abstract

Abstract: The aim of this study was to establish a multiplex PCR method for simultaneously detecting porcine reproductive and respiratory syndrome virus (PRRSV),porcine circovirus type 2 (PCV2),porcine pseudorabies virus (PRV) and Mycoplasma hyopneumoniae (Mh).Four pairs of primers were synthesized according to the reference.The multiplex PCR method was developed by optimizing the reaction condition,specificity and sensitivity detection.Four different amplicons with size of 424,490,298 and 360 bp for PRRSV,PCV2,PRV and Mh,respectively,were yielded.The sensitivity of multiplex PCR indicated that the detection limit was 3 copies by using Multiplex PCR Master Mix,and other common pathogens were not amplified.A total of 60 specimens from piglets were tested by multiplex PCR method.The positive accordance rate between simple and multiplex PCR was 100%.This study indicated that multiplex PCR might be a useful tool for rapid and sensitive etiological diagnosis and provided an effective technical support for pathogenic molecular epidemiology investigation.

Key words: porcine reproductive and respiratory syndrome virus (PRRSV); porcine circovirus type 2 (PCV2); porcine pseudorabies virus (PRV); Mycoplasma hyopneumoniae (Mh); multiplex PCR; detection

CLC Number:

S858.28

ZHANG Xun, WU Peng, LI Shi-zhen, FU Yin-feng, XU Zhui, LIANG Tian, YANG Su-fang, CHEN Xin-kai, QI Ya-yin, SHENG Jin-liang. Establishment and Preliminary Application of the Multiplex PCR Method for Detection of 4 Kinds of Swine Diseases[J]. , 2015, 42(10): 2612-2618.

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Establishment and Preliminary Application of the Multiplex PCR Method for Detection of 4 Kinds of Swine Diseases

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