实时荧光定量PCR技术在SPF鸡四种垂直传播疾病监测中的应用  

Application of SYBR Green Fluorescent Quantitative Real-Time PCR in the Detection of Four Vertical Transmission Viruses in SPF Chickens

作　　者：王静[1] 陈冬妮[2] 欧芷华[2] 袁文[1] 邓庆丽[2] 陆勇军[2] 张钰[1] 

机构地区：[1]广东省实验动物监测所(广东省重点实验室),广州510262 [2]中山大学生命科学学院,广州510275

出　　处：《中国比较医学杂志》2012年第8期6-14,共9页Chinese Journal of Comparative Medicine

基　　金：广东省教育部产学研结合项目(2008B090500205);广东省实验动物重点实验室(2007B060101002)联合资助

摘　　要：目的探讨荧光定量PCR检测技术对SPF鸡四种垂直传播病毒的检测应用。方法采集60份SPF鸡及70份普通鸡群蛋清、泄殖腔试子样品,提取样品核酸,分别进行ARV、REV、CAV、ALV四种病毒实时荧光定量PCR检测,根据标准曲线及溶解曲线分析判读样品病毒拷贝数。结果 SPF鸡ALV 2份阳性,检出率3.3%,其余病毒检测均为阴性;普通鸡样品REV检测2份阳性,检出率2.9%,ALV 10份阳性,检出率14.3%。结论荧光定量PCR检测方法最低可检测到100个拷贝核酸,检测灵敏度较高,有望应用于SPF鸡临床样品的病原检测。Objective To apply SYBR green I fluorescent quantitative real-time PCR（QPCR） in detecting 4 vertical transmission viruses of chicken and to evaluate its application value for the quality control of SPF chickens.Method Quantitative real-time PCR technique was used to detect REV,ALV,ARV and CAV RNA,DNA or provirus DNA in 30 cloacal swab samples and 30 eggs of SPF chicken samples as well as 40 cloacal swab samples and 30 eggs of common chicken samples.Standard curve and melting curve analyses were conducted to obtain the quantitative result.Results Out of the SPF chicken samples,2/30（3.3%） were ALV-positive,and the three other viruses were negative.For the common chicken samples,2/70（2.9%） were REV-positive,and 10/70（14.3%） were ALV-positive.Conclusions These QPCR assays show better sensitivity and specificity for detecting the 4 viruses in SPF chickens with a detection limit of 100 copies of target DNA per-reaction,and have a bright prospect in clinical application.

关 键 词：实时荧光定量PCR 禽网状内皮增生症病毒 鸡传染性贫血病毒 禽白血病病毒 禽呼肠孤病毒 SPF鸡 

分 类 号：R33[医药卫生—人体生理学]