山羊传染性胸膜肺炎荧光定量PCR检测方法的初步建立  

Development of the real-time PCR assay for detection of contagious caprine pleuropneumonia

作　　者：赵文华[1] 李富祥[1] 杨仕标[1] Zhao Wenhua;Li Fuxiang;Yang Shibiao(Yunnan Tropical and Subtropical Animal Viral Disease Laboratary,Yunnan Animal Science and Veterinary Institute,Yunnan Kunming 650224)

机构地区：[1]云南省畜牧兽医科学院云南省热带亚热带动物病毒病重点实验室,云南昆明650224

出　　处：《现代畜牧兽医》2022年第1期19-22,共4页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：国家自然科学基金项目(32060809);云南省重点新产品开发计划项目(2014BB014)。

摘　　要：为实现山羊传染性胸膜肺炎(CCPP)快速检测,研究基于病原体山羊支原体山羊肺炎亚种的16S rRNA基因,设计合成特异性引物及探针,采用5’FAM-TAMRA3’标记物进行标记。结果显示,试验建立的CCPP-16S实时荧光定量PCR方法可特异性检测CCPP,产生特异性荧光信号,对相关其他病菌无法检出荧光信号;以CCPP-16S载体质粒为标准品,构建荧光标准曲线,探针的检测敏感度可达10^(2.06)copies/μL DNA。研究表明,试验建立的实时荧光定量PCR方法可快速特异性检测CCPP,有待临床实践应用验证。To achieve rapid detection of contagious caprine pleuropneumonia(CCPP),the study was based on the 16 S rRNA gene of the pathogen Mycoplasma capricolum sub.Capripneumoniae of goat pneumonia,design and synthesis of specific primers and probes,used 5’FAM-TAMRA3’label for labeling.The results showed that the CCPP-16 S realtime PCR method established by the institute can specifically detected CCPP and generate specific fluorescent signals,but it cannot detect fluorescent signals for other related bacteria.The CCPP-16 S vector plasmid was used as the standard to construct a real-time standard curve,and the detection sensitivity of the probe could reach 10^(2.06) copies/μL DNA.The experiment indicates that the real-time PCR method established in the experiment can quickly and specifically detected CCPP,which need to be verified in clinical practice.

关 键 词：山羊传染性胸膜肺炎 支原体山羊肺炎亚种 16S rRNA基因 实时荧光定量PCR 探针 

分 类 号：S855[农业科学—临床兽医学]