摘要
为建立一种能够同时检测山羊支原体山羊肺炎亚种和多杀性巴氏杆菌的双重PCR方法,本研究采用2对特异性引物,对退火温度和引物浓度比进行了优化,成功建立了一种能同时检测上述两种病原的双重PCR方法。结果显示,该方法具有很好的特异性,仅对山羊支原体山羊肺炎亚种和多杀性巴氏杆菌有扩增,而对其他常见的羊呼吸道病原无扩增;该方法对两种病原的检测限分别为32pg和50pg,与单独PCR相同;26份临床样品中山羊支原体山羊肺炎亚种的检出率为23.1%,多杀性巴氏杆菌的检出率为26.9%。所建立的双重PCR具有特异性好、灵敏度高的特点,为临床上这两种病原感染的快速诊断和流行病学调查等提供了更为有用的手段。
The purpose of this study was to establish a duplex PCR method for simultaneous detection of Mycoplasma capricolum subsp.capripneumoniae （Mccp）and Pasteurella multocida .Two sets of primers specific to Mccp and P .multocida were applied for development of the duplex PCR.After optimization of PCR components and reaction profile,a duplex PCR method was established.The results demonstrated that the assay is highly specific to Mccp and P .multocida ,and no-target pathogens were not detected. The detection limits of the assay were determined to be 32 pg for Mccp DNA and 50 pg for P .multocida DNA,respectively,which was as sensitive as single PCR methods.The developed duplex PCR could de-tect both of the pathogens from clinical samples.The duplex PCR established in this study will be useful for clinical detection,identification and epidemiological investigation of Mccp and P .multocida.
出处
《动物医学进展》
CSCD
北大核心
2014年第12期10-13,共4页
Progress In Veterinary Medicine
基金
西南民族大学大学生创新创业训练计划(201310656013)
