猪附红细胞体快速诊断方法的研究

Establishment of PCR Assay for detecting Mycoplasma suis(Eperythrozoon suis)

根据GenBank上已报道的猪附红细胞体16S rRNA的序列设计一对特异性引物,进行PCR扩增并将产物克隆到T-vector后测序.结果表明扩增到的目的基因片段为404 bp,与GenBank上Mycoplasma suis序列同源性为97%.试验建立的PCR诊断方法特异性强,与健康猪血液、猪链球菌、副猪嗜血杆菌、多杀性巴氏杆菌、胸膜肺炎放线杆菌、弓形虫、猪圆环病毒DNA无交叉反应,能检测出的最低DNA浓度是8.6 fg?μL-1.该方法具有快速、准确、敏感等特点,为浙江地区猪附红细胞体病的诊断及分子流行病学的调查提供了新的手段.

To establish a PCR-based （polymerase chain reaction） method for the detection of Mycoplasma suis （M. suis） which was named Eperythrozoon suis, a pair of primers were designed according to the published sequence data of the 16S ribosomal RNA gene of M. .suis. PCR product was cloned and sequenced. Sequence analysis showed that the gene fragment of 404 bp was 97% identical to the published data. No cross-reactivity was detected from normal swine blood, Streptococcus .suis, Hemophiluspara sui.s, pasteurellamultocida, Actinobacillus pleuropneumoniae, Toxoplasma gondii, PCV-2, and the lowest DNA concentration of M. suis detected was 8.6 fg·μL^-1 , This specific, sensitive and rapid PCR assay was suitable laboratory method for the detection of M. suis. It could also be used in research of epidemiology and early clinical diagnosis of M. suis in molecular level.