中国兽医科学2025，Vol.55Issue(9)：1182-1189,8.DOI:10.16656/j.issn.1673-4696.2025.0123
羊支原体肺炎病原三重PCR检测方法的建立
Establishment of a triple PCR detection method for ovine mycoplasmal pneumonia
摘要
Abstract
Ovine mycoplasmal pneumonia is a collective term for a category of diseases caused by various mycoplasmas.The pathogens reported in China primarily includ Mycoplasma ovipneumoniae(Mo),Mycoplasma capricolum subsp.Capri pneumoniae(Mccp)and Mycoplasma mycoides subsp.Capricolum(Mmc)and others.Genome collinearity analysis was used to establish a method that can quickly identify Mo,Mccp and Mmc.And a total of 87 primers were designed for detection.A triple PCR detection method for Mo,Mccp,and Mmc was established using selected primers,with specificity and sensitivity tests,as well as clinical sample detection,conducted post-optimization.The results showed that a multiplex PCR detec-tion method was established by using the most suitable combination of screened primers,which could specifically amplify three target bands:Mo(310 bp),Mccp(668 bp)and Mmc(310 bp).And the optimal elon-gation time and annealing temperature were 10 s and 56℃.There was no cross-reaction with other pathogens(Mycobacterium bovis,Brucella,Pasteurella multocida,Staphylococcus aureus,Salmonella).The lowest detection copy numbers for the constructed positive standard plasmids were 102 copies/μL,101 copies/μL,and 102 copies/μL,respectively.The results showed that the multiplex PCR method estab-lished could be used for rapid differential diagnosis and epidemiological investigation of Mo,Mccp and Mmc infections,and had the characteristics of clinical convenience and rapid diagnosis.关键词
羊支原体肺炎/多重PCR/绵羊肺炎支原体/山羊支原体山羊肺炎亚种/丝状支原体山羊亚种Key words
ovine mycoplasmal pneumonia/multiplex PCR/Mo/Mccp/Mmc分类
农业科技引用本文复制引用
吕皓晴,刘颖,张粟子,刘立元,颜新敏,郝华芳,储岳峰,许健,袁律峰..羊支原体肺炎病原三重PCR检测方法的建立[J].中国兽医科学,2025,55(9):1182-1189,8.基金项目
国家重点研发计划项目子课题(2022YFD180070202) （2022YFD180070202）
中国农业科学院创新工程项目(CAAS-ASTIP-2021-LVRI) （CAAS-ASTIP-2021-LVRI）
甘肃省科技重大专项(23ZDNA007) （23ZDNA007）
