鸡滑液囊支原体和禽呼肠病毒的实时荧光定量PCR快速检测  

Rapid Detection of Mycoplasma synoviae and Avian Reovirus by Real-time PCR

作　　者：黄莉 谢芝勋 王盛 邓显文 谢志勤 谢丽基 曾婷婷 黄娇玲 张艳芳 

机构地区：[1]广西壮族自治区兽医研究所/广西兽医生物技术重点实验室,广西南宁530001

出　　处：《动物医学进展》2018年第2期1-6,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31660715);广西科技厅重点研发计划项目(桂科AB16380003);广西自然科学基金项目(2014GXNSFCA118006);广西壮族自治区直属科研院所基本业务费项目(桂科专15-1);广西兽医生物技术室开放基金项目(14-045-31-B-1;12-071-28-B-2);国家"万人计划"领军人材专项项目(2016-37)

摘　　要：根据禽呼肠病毒σC和鸡滑液囊支原体vlhA基因的保守序列,分别设计了特异性引物和不同荧光基团标记的TaqMan探针,优化反应条件,建立能同时鉴别诊断鸡滑液囊支原体和禽呼肠病毒(ARV)的检测方法。结果表明,该方法能够特异地鉴别检测ARV和MS,与其他病原体无交叉反应,检测敏感性均可达到2×10拷贝数。此外,该方法抗干扰能力强,具有良好而稳定的准确性,临床样品检出率与传统检测方法相符。因此,该基于TaqMan探针建立的荧光定量PCR具有特异、灵敏、准确、实时、重复性好等优点,不仅可用于鸡群中这两种病原体的鉴别检测和疫病监测,也有利于这两种病原体感染的有效防控。Based on highly conserved regions in the ARV and MS genomes, the specific primers and probeswere designed,and the reaction conditions were optimized, and the duplex real-time PCR assay was devel-oped for the rapid detection and quantification of ARV and MS. The results showed that the duplex real-time PCR assay was capable of ARV- and MS-specific detection without cross-reaction with other non-tar-geted avian pathogens. The sensitivity of this assay was 2 X 101 copies for a recombinant plasmid containingARV aC or MS vlhA gene. This new assay was also reproducible and stable. All tested field samples ofARV and/or MS were detectable with this duplex real-time PCR assay compared with pathogen isolationand identification as well as serological tests. This duplex real-time PCR assay is highly specific, sensitiveand reproducible and thus could provide a rapid,specific and sensitive diagnostic tool for the simultaneousdetection of ARV and MS in poultry flocks. The assay will be useful not only for clinical diagnostics anddisease surveillance but also for the efficient control and prevention of ARV and MS infections.

关 键 词：荧光定量PCR TAQMAN 鸡滑液囊支原体 禽呼肠病毒 

分 类 号：S858.31[农业科学—临床兽医学]