华南虎源血巴尔通体的PCR检测

PCR Amplification of Haemobartonella of South China Tigers

为了快速准确检测华南虎感染的血巴尔通体种类,我们根据已发表的猫血巴尔通体的16S rRNA基因序列,利用软件Primer Premier 5.0设计2对特异性引物,以华南虎源血巴尔通体基因组DNA进行PCR检测。2对引物分别扩增出1452 bp和170 bp的rDNA序列,1452 bp序列与GenBank^(TM)发表的3种猫血巴尔通体序列相似性分别为99.3%、91.7%和79.5%,而170 bp序列与序列AM748929的相似性为100%。实验结果表明华南虎感染的血巴尔通体为大型猫血巴尔通体,实验建立的PCR检测方法为华南虎血巴尔通体病的诊断及分子流行病学的调查提供了新的手段。

Haemobartonella infecting South China tigers（Panthera tigris amoyensis） was detected by PCR amplification of 2 pairs of primers,designed according to the published base sequence of 16S rRNA gene of Haemobartonella felis of the domestic cat using software Primer Premier 5.0,were amplified and 2 fragments,1452 and 170 respectively were obtained and se-quenced. The sequence of 1452 bp were compared to homologous sequences of Candidatus mycoplasma Haemofelis,Candidatus mycoplasma Haemominuium and Candidatus mycoplsma turicensi in GenBank^（TM） and the homology of sequences were 99.3%, 91.7%and 79.5%respectively.The sequence of 170 bp fragment was compared to AM748929 and the homology was 100%. Results revealed that Haemobartonella infecting the tiger was a member of Candidatus mycoplasma Haemofelis and these primers could be used to PCR detection and diagnosis of Haemobartonella of South China tigers.