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当前位置: 首页 > 文章 > 山羊传染性胸膜肺炎荧光定量PCR检测方法的初步建立 现代畜牧兽医 2022 (1) 19-22
Position: Home > Articles > Development of the real-time PCR assay for detection of contagious caprine pleuropneumonia Modern Journal of Animal Husbandry and Veterinary Medicine 2022 (1) 19-22

山羊传染性胸膜肺炎荧光定量PCR检测方法的初步建立

作  者：
赵文华;李富祥;杨仕标
单  位：
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
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
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102.06copies/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。
译  名：
Development of the real-time PCR assay for detection of contagious caprine pleuropneumonia
作  者：
Zhao Wenhua;Li Fuxiang;Yang Shibiao;Yunnan Tropical and Subtropical Animal Viral Disease Laboratary,Yunnan Animal Science and Veterinary Institute;
关键词：
Contagious caprine pleuropneumonia;;Mycoplasma capricolum sub.Capripneumoniae;;16S rRNA gene;;Real-time PCR;;Probe
摘  要：
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 102.06 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.

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