单 位：
浙江出入境检验检疫局;浙江大学动物预防医学研究所
关键词：
牛附红细胞体;聚合酶链式反应(PCR);克隆
摘 要：
根据已报道的牛附红细胞体16S rRNA的序列设计1对特异性引物,进行PCR。将PCR产物克隆并测序,结果显示所得基因片段为667 bp,与GenBank上Mycoplasma wenyonii序列同源性达到98.83%。试验建立的PCR诊断方法特异性强,与健康牛血液、牛无乳链球菌、金黄葡萄球菌、多杀性巴氏杆菌、胸膜肺炎放线杆菌、大肠杆菌的DNA无交叉反应,能检测的牛附红细胞体最低DNA质量浓度是13.6 ng/L。应用该方法对130个奶牛血样进行牛附红细胞体病检测,结果表明总阳性感染率为24.62%。该方法具有快速、准确、敏感等特点,为牛附红细胞体病的诊断及分子流行病学的调查提供新的手段。
译 名：
Establishment of a diagnostic PCR assay for Mycoplasma wenyonii
作 者：
HOU Yu-hui1,WU Sheng-li1,2,CAI Wei-ming3,ZHAO Xian-feng1,DU Ai-fang1*(1.Institute of Preventive Veterinary Medicine,Zhejiang University,Hangzhou 310029,China;2.Jinhua Entry-exit Inspection and Quarantine Bureau,Jinhua,Zhejiang 321001,China;3.Zhejiang Entry-exit Inspection and Quarantine Bureau,Hangzhou 310004,China)
关键词：
M.wenyonii;PCR;clone
摘 要：
To establish a PCR-based(polymerase chain reaction) method for the detection of Mycoplasma wenyonii,a pair of primers was designed according to the published sequence data of the 16S ribosomal RNA gene of M.wenyonii.PCR product was cloned and sequenced.Sequence analysis showed that the gene fragment obtained was 667 bp and was 98.83% identical to the published data.The special and sensitive tests showed that no PCR product was detected from normal cattle blood,Streptococcus agalactiae,Staphylococcus aureus,Pasteurella multocida,Actinobacillus pleuropneumoniae,Escherichia coli,and the lowest DNA concentration of M.wenyonii that could be detected was 13.6 ng/L.Applying the established PCR,24.62% out of 130 samples from different dairy farms were found infected with M.wenyonii.This specific,sensitive and rapid PCR assay was suitable for laboratory detection of M.wenyonii.It could also be used for research of epidemiology and early clinical diagnosis of M.wenyonii in molecular level.
