副猪嗜血杆菌PCR检测方法的建立与应用  

Establishment and application of a PCR method for detection of Haemophilus parasuis infection

作　　者：马超锋 余洪涛 彭燕 左春生[3] 王俊锋[3,4] 

机构地区：[1]信阳市动物疫病预防控制中心,河南信阳464000 [2]淮滨县畜产品检验检测中心,河南信阳464400 [3]信阳农林学院牧医工程学院,河南信阳464001 [4]信阳市动物健康养殖营养工程技术研究中心,河南信阳464001

出　　处：《畜牧与兽医》2017年第2期89-93,共5页Animal Husbandry & Veterinary Medicine

基　　金：信阳市科技攻关项目

摘　　要：为了快速检测临床上副猪嗜血杆菌(Haemophilus parasuis,Hps)感染,根据Gen Bank中Hps的16S rRNA基因保守区序列,设计1对特异性引物,利用PCR方法检测Hps,并进行特异性、敏感性及临床检测。结果显示:PCR反应选择58℃为最适退火温度,0.5μL(20μmol/L)为最适引物量,模板浓度为7.2×10^(-6)μg/mL时即可扩增出814 bp特异性目的片段,与细菌分离结果一致。结果表明:该方法敏感、快速、特异,可用于临床上Hps感染的快速检测。To establish a rapid method for detection of Haemophilus parasuis,PCR was developed with a pair of primers,which was designed according to the conserved sequence of 16 S rRNA gene of H.parasuis available in Gen Bank.The results showed that the optimal reaction system is as follows：the optimum annealing temperature is 58 ℃;the optimal concentration of primers is 0.5 μL（20 μmol/L）;the concentration of the template is 2×10^（-6）μg/mL.The specific fragment of 814 bp could be amplified from both the reference and clinically isolated strains by this PCR method.Meanwhile,the target fragment could be amplified from the samples of the pigs with suspected clinical signs by this established PCR method,and the result was consistent with the bacteria isolation.The results demonstrated that the PCR method was sensitive,specific and could be used for further clinical application.

关 键 词：副猪嗜血杆菌 PCR 特异性 敏感性 

分 类 号：S852.6[农业科学—基础兽医学]