动物医学进展Issue(10)：1-6,6.
禽肺炎病毒实时荧光定量 RT-PCR 检测方法的建立
Development of Real-time RT-PCR for Detection of Avian Pneumovirus in Chickens
摘要
Abstract
A SYBR Green Ⅰ real-time RT-PCR was developed and applied to detect avian pneumovirus subtype C.A pair of specific primers were designed and synthesized according to the fusion gene of avian pneumovirus subtype C in GenBank.The reaction parameters,such as the concentration of primers,and the reaction buffer,were optimized to develop a SYBR Green Ⅰ real-time RT-PCR for the rapid detection of avian pneumovirus subtype C.The sensitivity and specificity were tested by this method.The results in-dicated that only avian pneumovirus strain had the positive curve in this assay.As limit as 10 copies of plas-mid DNA isolated from competent cells that transferred avian pneumovirus gene was detected in this test. 54 of clinical samples collected from chicken lungs and trachea were detected by this method.It showed that all of clinical samples were negative for avian pneumovirus subtype C.It indicated that this SYBR Green Ⅰ real-time RT-PCR was a good method for detection of avian pneumovirus.It was a quick,sensi-tive,specific and quantitative method for identification of avian pneumovirus in chickens in the future.关键词
SYBR Green Ⅰ/实时荧光定量 RT-PCR/检测/禽肺炎病毒Key words
SYBR Green Ⅰ/real-time RT-PCR/detection/Avian pneumovirus分类
农业科技引用本文复制引用
谢志勤,谢芝勋,刘加波,庞耀珊,邓显文,谢丽基,范晴,罗思思..禽肺炎病毒实时荧光定量 RT-PCR 检测方法的建立[J].动物医学进展,2014,(10):1-6,6.基金项目
广西重大专项项目（桂科重1222003-2）；广西特聘专家专项项目 （）
