摘要
为建立一种特异、敏感的绵羊肺炎支原体PCR检测技术,根据绵羊肺炎支原体p113基因设计引物,并对其特异性进行了评价,同时,对所建立的PCR方法的敏感性和临床样本中绵羊肺炎支原体的检出率进行了分析,并与现有的基于16SrRNA的PCR方法进行了比较。结果显示,该方法能特异性扩增绵羊肺炎支原体参考菌株Y-98及临床分离株,对其他羊的致病性支原体和无关病原不检出;检测灵敏性为44fg,为现有的基于16SrRNA基因的PCR技术的1 000倍;对临床采集的肺组织样本和鼻腔拭子中绵羊肺炎支原体的检出率明显高于基于16SrRNA的PCR方法。研究结果表明,所建立的基于p113基因的PCR技术具有特异、敏感等特点,为绵羊肺炎支原体的检测、鉴定以及流行病学调查提供了有用的工具。
The aim of this study was to establish a specific and sensitive PCR assay for detection of Mycoplasrna ovipneumoniae. A pair of primers targeting p113 gene of M. ovipneumoniae was designed and the specificity of the primers was evaluated. The sensitivity and detection rates for clinical samples were also investigated and compared to existing 16 S rRNA-based PCR. The results showed that the PCR assay developed in this study could detect both the reference strain Y-98 and other field isolates,while no positive amplification was observed from other pathogenic Mycoplasma species and bacteria in sheep and goats. The detection limit of the assay was determined to be 44 fg,1 000 times more sensitive than 16 S rRNA-based PCR. It was also demonstrated that,compared with 16 S rRNA-based PCR,the p113-based PCR had signi- ficantly higher detection rates for both lung tissues and nasal swabs. These results suggest that the deve- loped PCR assay is specific and sensitive,and will be useful for detection,identification and epidemiological investigation of M. ovipneumoniae.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第11期1157-1161,共5页
Chinese Veterinary Science
基金
国家自然科学基金项目(31272588)
西南民族大学研究生创新型科研项目(CX2013SZ70)
