摘要
为建立同时检测绵羊肺炎支原体(MO)和溶血性曼氏杆菌(Mh)的方法,本研究根据MO的hsp70基因和Mh的gcp基因分别设计引物,建立了同时检测这两种病原的双重PCR检测方法。实验结果显示该方法能够特异性的扩增MO(135 bp)和Mh(227 bp)DNA片段,其最低检出量分别为2.06×103拷贝/μL和6.37 cfu/mL,与单一PCR相同。该方法对常见的致病菌无交叉反应。对临床样本的检测结果显示MO和Mh的检出率分别为56.25%和52.50%,与单独PCR符合率为100%。研究结果表明,本研究所建立的双重PCR检测方法具有特异性强、敏感性高等特点,为临床上MO和Mh混合感染的快速检测、鉴定以及流行病学调查提供了方便、快捷、准确的方法。
The present study reports the development of a duplex PCR method for simultaneous detection of Mycoplasma ovipneumoniae （MO） and Mannheimia haemolytica （Mh） by using primers targeting hsp70 gene of MO and gcp gene of Mh,respectively.The detection results showed the established duplex PCR assay were able to specific to amplify the DNA fragments of 135 bp from MO and 227 bp from Mh,but no amplification was found from the other related pathogenic bacteria.While,the detection limits of the assay were 2.06×103 copies/μL for MO and 6.37 cfu/μL for Mh,respectively,which were identical to the sensitivity of single PCR assays.Detections of 160 clinical samples by the duplex PCR showed the positive rates were 56.25% for MO and 52.50% for Mh.These results suggested that the established duplex PCR assay is specific and sensitive,which has a potential to be applied for detections and epidemiological investigation of MO and Mh infections.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2014年第10期788-791,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
四川省杰出青年学术技术带头人培育计划(2013JQ0040)
