猪多杀性巴氏杆菌和副猪嗜血杆菌双重及多重PCR检测方法的建立  

Establishment of double and multiple PCR detection methods for Pasteurella multocida and Haemophilus parasuis

作　　者：朱新贵 田亚平[2] 王亚珊 吴艳芳 刘勇军[2] 姚强[2] 姜艳彬[2] 张锐利[1] 侯东军[2] ZHU Xingui;TIAN Yaping;WANG Yashan;WU Yanfang;LIU Yongjun;YAO Qiang;JIANG Yanbin;ZHANG Ruili;HOU Dongjun(College of Life Science, Tarim University, Alar 843300, China;China Animal Disease Control Centre, Beijing 100125, China;Heze Vocational Institute. Heze 274031. China)

机构地区：[1]塔里木大学生命科学学院,新疆阿拉尔843300 [2]中国动物疫病预防控制中心,北京100125 [3]菏泽职业学院,山东菏泽274031

出　　处：《黑龙江畜牧兽医》2018年第8期75-78,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：“十二五”国家科技支撑计划项目(2014BAD13B05)

摘　　要：为了准确检测临床上猪多杀性巴氏杆菌（Pasteurella multocida）和副猪嗜血杆菌（Haemophilus parasuis）的感染,根据Gen Bank中副猪嗜血杆菌16S rRNA、PILA、LIPO基因保守序列和猪多杀性巴氏杆菌的16S rRNA、KMT1基因保守序列建立双重及多重PCR检测方法,通过对其退火温度的筛查,确定最优退火温度,然后进行方法的验证。结果表明：与传统方法相比,所建立的双重及多重PCR检测方法具有快速、准确度高、特异性强的特点。说明本方法可以用于猪多杀性巴氏杆菌和副猪嗜血杆菌在临床上的快速检测。To detect Pasteurella multocida and Haemophilus parasuis infection,set of double and multiple PCR methods were established based on the conservative gene sequences from Gen Bank of 16 S rRNA,PILA and LIPO in Haemophilus parasuis and conservative gene sequences of16 S rRNA,KMT1 in Pasteurella multocida. Then,the optimum annealing temperature was determined by screening the annealing temperature.Finally,the method was verified. Results showed that,compared with the traditional method,the established double and multiple PCR methods have the characteristics of high speed,high accuracy and strong specificity,which can provide a new technique for the rapid clinical detection of Pasteurella multocida and Haemophilus parasuis.

关 键 词：猪多杀性巴氏杆菌 副猪嗜血杆菌 多重PCR 16S RRNA基因 外毒素基因 

分 类 号：S852.61[农业科学—基础兽医学] Q781[农业科学—兽医学]