绵羊附红细胞体感染PCR检测方法的建立

Establishment of the PCR Method for Detection of Ovine Eperythrozoonsis

为建立特异、敏感、快速的绵羊附红细胞体感染诊断方法,本试验根据已测得的绵羊附红细胞体16S rRNA基因序列(Gene Bank:EU916726、FJ440328),设计1对特异性引物,建立了绵羊附红细胞体的PCR诊断方法。特异性实验和敏感性实验结果表明,该方法与绵羊肺炎支原体、大肠杆菌、葡萄球菌、白色念珠菌、枯草杆菌均无交叉反应,能检测到绵羊附红细胞体最低DNA量为5.2pg/μL,在-80℃存放1年的血样中也检测到阳性样品。通过临床血样检测,证明该方法可以用于本病的早期感染和隐性感染的检测。

In order to establish a specific, sensitive, fast detection assay for Mycoplasma ovis infection, this study designed a pair genus specific primers based on the 16SrRNA gene（GeneBank：EU916726, FJ440328） of E. ovis sequenced. The assay was able to detect 5.2pg target DNA and had no cross reaction with Monilia albican, E. coli, Bacillus Subtilis, Streptococcus ,Staphylococcus ,Mycoplasma ovipneumoniae. Besides, it can also detect the blood sample stored at -80℃ more than twelve months. The clinic blood sample detections have proved that the assay could be used in E. ovis infection.