CAJ | 학술논문

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32 pg 和50 pg，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。
위건립일충능구동시검측산양지원체산양폐염아충화다살성파씨간균적쌍중 PCR 방법，본연구채용2대특이성인물，대퇴화온도화인물농도비진행료우화，성공건립료일충능동시검측상술량충병원적쌍중 PCR 방법。결과현시，해방법구유흔호적특이성，부대산양지원체산양폐염아충화다살성파씨간균유확증，이대기타상견적양호흡도병원무확증；해방법대량충병원적검측한분별위32 pg 화50 pg，여단독 PCR상동；26빈림상양품중산양지원체산양폐염아충적검출솔위23．1％，다살성파씨간균적검출솔위26．9％。소건립적쌍중 PCR 구유특이성호、령민도고적특점，위림상상저량충병원감염적쾌속진단화류행병학조사등제공료경위유용적수단。
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.