PRRSV和PCV-2二重PCR检测方法的建立及初步应用  

Establishment and Application of Duplex PCR for PRRSV and PCV-2 Detection

作　　者：王林[1,2] 吴发兴[2] 吴美芹[3] 高许雷[1,2] 李平[2,4] 朱紫祥[2,5] 张燕霞[2] 张志[2] 李晓成[2] 单虎[1] 

机构地区：[1]青岛农业大学,山东青岛266109 [2]中国动物卫生与流行病学中心,山东青岛266032 [3]招远市畜牧局,山东烟台265400 [4]新疆农业大学,新疆乌鲁木齐830052 [5]西北农林科技大学,陕西杨陵712100

出　　处：《动物医学进展》2010年第6期1-5,共5页Progress In Veterinary Medicine

基　　金：十一五科技支撑计划项目(2007BAD86B04)

摘　　要：参考GenBank上已发表的猪繁殖与呼吸综合征病毒Nsp2基因序列和猪圆环病毒2型ORF1基因序列,分别设计并合成了可用于检测猪繁殖与呼吸综合征病毒和猪圆环病毒2型的2对引物,扩增目的片段大小分别为421 bp和714 bp。在建立了用于检测猪繁殖与呼吸综合征病毒和猪圆环病毒2型的单项PCR检测方法并优化单项PCR条件的基础上,建立了针对两种病毒的二重PCR检测方法。通过检测143份临床病料对二重PCR方法和单项PCR/RT-PCR方法进行了对比验证。结果显示,两者的总符合率在92.6%以上,表明该二重PCR检测方法可以用于临床病料的检测。Two pairs of specific primers were designed according to the sequences of PRRSV Nsp2 gene and PCV-20RF1 gene availiable on GenBank respectively. The amplified fragment of PRRSV was 421bp,and 714： bp for PCV-2. Two PCR/RT-PCR methods for detecting PRRSV and PCV-2 were developed using the two pairs of primers. Based on the two established PCR methods above,the duplex PCR method for detecting both PRRSV and PCV-2 was developed through combination and optimization of two methods. 143 clinical samples were detected by both this duplex PCR method and two seperate PCR/RT-PCR methods. The results showed 92.6% consistency between the duplex PCR method and two seperate PCR/RT-PCR methods,which suggested that the established duplex PCR method can be used in clinical detection for PRRSV and PCV-2.

关 键 词：猪繁殖与呼吸综合征病毒 猪圆环病毒2型 二重PCR 

分 类 号：S852.659.6[农业科学—基础兽医学] S852.659.2[农业科学—兽医学]