牛无浆体和山羊无浆体双重PCR检测方法的建立  

Development of duplex PCR for simultaneous detection of Anaplasma bovis and Anaplasma capra

作　　者：彭永帅[1] 张利卫 王军[1] 李月勤[1] 王海燕 PENG Yongshuai;ZHANG Liwei;WANG Jun;LI Yueqin;WANG Haiyan(College of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou 450064,China)

机构地区：[1]河南牧业经济学院动物医药学院,河南郑州450046

出　　处：《畜牧与兽医》2020年第7期102-107,共6页Animal Husbandry & Veterinary Medicine

基　　金：河南牧业经济学院博士科研启动基金(53000183)。

摘　　要：无浆体是一种专性血液寄生的可经硬蜱传播的病原体,世界范围内分布广泛。本研究旨在建立一种可以同时检测牛无浆体和山羊无浆体的双重PCR方法。通过对2种病原gltA基因和msp4基因位点引物的筛选,PCR反应体系中引物剂量、酶剂量和PCR反应条件中的退火温度、循环次数的优化,最终确定了一个引物组合和最适的PCR反应体系和条件,并对其进行了重复性和特异性验证。结果表明,本研究建立的双重PCR方法有效,操作简便、特异性、重复性良好,为兽医工作者和科研人员提供了一种山羊无浆体和牛无浆体感染快速鉴别诊断和流行病学监测工具。Anaplasmais an obligate blood parasite that can be transmitted by ticks and is widely distributed in the world.This study wasaimed to establish a duplex PCR method for simultaneous detection ofA.bovis andA.capra.Primers against the conserved regions of theA.bovis gltA andA.capra groELgenes were designed.A range of PCR-related parameters were evaluated,such as the dosage of primers,enzyme,annealing temperature,etc.So as to finally determine a primer combination and the most appropriate PCR reaction system and conditions,which were verified for specificity,sensitivity and repeatability.The results showed that the duplex PCR method established in this study was effective,specific and repeatable,and served to be a very efficient tool for veterinary workers and scientific researchers in rapid differential diagnosis and epidemiological monitoring ofA.bovis andA.capra.

关 键 词：二重PCR 山羊无浆体 牛无浆体 

分 类 号：S855.9[农业科学—临床兽医学] S858.23[农业科学—兽医学]