羊附红细胞体巢式PCR检测方法的建立及临床应用  

Establishment of a nested PCR assay for detection of Mycoplasma ovis

作　　者：赵晓薇[1] 巴彩凤[1] 苏玉虹[2] 苏荣健[3] 冯会权[1] 

机构地区：[1]辽宁医学院实验动物中心,辽宁锦州121001 [2]辽宁医学院动物科学技术学院,辽宁锦州121001 [3]辽宁医学院科学实验中心,辽宁锦州121001

出　　处：《中国预防兽医学报》2008年第7期552-556,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：辽宁省科技基金资助项目(2006408002-4)

摘　　要：根据GenBank上发表的羊附红细胞体16SrRNA基因序列(登录号AF338268)设计2对引物,用巢式PCR方法扩增16SrRNA的部分序列,将目的片段克隆并测序。测序结果与AF338268相似性达99%以上,只有3bp的差异。该方法与猪附红细胞体、羊肺炎支原体、链球菌、葡萄球菌和大肠杆菌均无交叉反应,能检测到的最低DNA量为25fg。用于检测的28份临床样本中,23份为阳性。所建立的巢式PCR检测方法具有较高的敏感性和特异性,可用于羊附红细胞体病急性感染和隐性感染的早期诊断,为该病的临床检测、流行病学调查、进出口检疫和实验室研究提供了新的技术手段。A nested PCR assay was developed using two pairs of primers derived from the 16S rRNA gene of Mycoplasma ovis, The assay was able to detect 25 fg target RNA and had no cross reaction with Mycoplasma suis, Mycoplasma ovipneumoniae, Streptococcus, Staphylococci and E.coli. Tests on blood samples from 28 borderline cases and 23 samples were shown positive. These results suggested that the nested PCR assay was a sensitive and effective method which could be used for clinical diagnose, epidemiologic and other applications.

关 键 词：羊附红细胞体 羊附红细胞体病 巢式PCR 

分 类 号：S855.99[农业科学—临床兽医学]