建立 O 抗原基因簇特异性 PCR 方法用于快速分型中国六群流行的钩端螺旋体。
Development of O-antigen gene cluster-specific PCRs for rapid typing six epidemic serogroups of Leptospira in China.
机构信息
Department of Medical Microbiology and Parasitology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
出版信息
BMC Microbiol. 2010 Mar 3;10:67. doi: 10.1186/1471-2180-10-67.
BACKGROUND
Leptospira is the causative agent of leptospirosis. The O-antigen is the distal part of the lipopolysaccharide, which is a key component of outer membrane of Gram-negative bacteria and confers serological specificity. The epidemiology and clinical characteristics of leptospirosis are relative to the serology based taxonomic unit. Identification of Leptospira strains by serotyping is laborious and has several drawbacks.
RESULTS
In this study, the O-antigen gene clusters of four epidemic Leptospira serogroups (serogroup Canicola, Autumnalis, Grippotyphosa and Hebdomadis) in China were sequenced and all genes were predicted in silico. Adding published sequences of two serogroups, Icterohaemorrhagiae (strain Lai and Fiocruz L1-130) and Sejroe (strain JB197 and L550), we identified six O-antigen-specific genes for six epidemic serogroups in China. PCR assays using these genes were developed and tested on 75 reference strains and 40 clinical isolates.
CONCLUSION
The results show that the PCR-based assays can be reliable and alternative means for rapid typing of these six serogroups of Leptospira.
背景
钩端螺旋体是钩端螺旋体病的病原体。O-抗原是脂多糖的远端部分，是革兰氏阴性细菌外膜的关键组成部分，赋予血清学特异性。钩端螺旋体病的流行病学和临床特征与基于血清学的分类单位有关。通过血清分型鉴定钩端螺旋体菌株既繁琐又有几个缺点。
结果
在这项研究中，对中国四个流行的钩端螺旋体血清群（血清群堪地拉、秋沙、格里波托萨和七日热）的 O-抗原基因簇进行了测序，并对所有基因进行了计算机预测。加入两个血清群（株 Lai 和 Fiocruz L1-130 的 Icterohaemorrhagiae 以及 JB197 和 L550 的 Sejroe）的已发表序列，我们确定了中国六个流行血清群的六个 O-抗原特异性基因。使用这些基因开发了 PCR 检测方法，并在 75 个参考菌株和 40 个临床分离株上进行了测试。
结论
结果表明，基于 PCR 的检测方法可以作为这些六种钩端螺旋体血清群快速分型的可靠替代方法。
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