评价用于检测蜱虫中伯氏疏螺旋体属的分子方法。

Evaluation of molecular methods for detection of Borrelia burgdorferi senso lato in ticks.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, P. R. China.

出版信息

Diagn Microbiol Infect Dis. 2012 May;73(1):80-3. doi: 10.1016/j.diagmicrobio.2012.02.016. Epub 2012 Apr 10.

DOI:10.1016/j.diagmicrobio.2012.02.016

PMID:22494560

Abstract

Borrelia burgdorferi sensu lato (s. l.), the agent of Lyme disease, is distributed widely worldwide. A large number of polymerase chain reaction (PCR) methods have been developed and used for detection of B. burgdorferi s. l. However, there is a lack of a reference standard because of the genetic diversity of the B. burgdorferi s. l. complex. In this study, 4 PCR methods, based on the OspA, flagellin, rrs, and P66 genes, for detection of B. burgdorferi s. l. were evaluated by detection of genomic DNA from 3 reference genospecies and tick samples. The sensitivity of the PCR methods was analyzed using serially diluted gDNA from B. afzelii (Bo23), B. burgdorferi sensu stricto (B31), and B. garinii (PBi). The performance of the PCRs was evaluated by detection of the gDNA of 543 ticks. The results showed that the PCRs targeting the OspA gene, fla gene, rrs gene, and P66 gene detected 37 (6.8%), 74 (13.6%), 16 (2.9%), and 14 (2.6%) tick samples, respectively. The PCR targeting the fla gene was the most sensitive method for the detection of B. burgdorferi s. l.

摘要

伯氏疏螺旋体（Borrelia burgdorferi），莱姆病的病原体，广泛分布于世界各地。已经开发并使用了大量聚合酶链反应（PCR）方法来检测伯氏疏螺旋体（Borrelia burgdorferi）。然而，由于伯氏疏螺旋体（Borrelia burgdorferi）复合体的遗传多样性，缺乏参考标准。在这项研究中，评估了基于 OspA、鞭毛蛋白、rrs 和 P66 基因的 4 种 PCR 方法，用于检测 3 种参考种属和蜱样本的基因组 DNA。通过对 Bo23、B31 和 PBi 三种血清型的 gDNA 进行连续稀释，分析了 PCR 方法的灵敏度。通过检测 543 只蜱的 gDNA 来评估 PCR 的性能。结果表明，针对 OspA 基因、fla 基因、rrs 基因和 P66 基因的 PCR 分别检测到 37（6.8%）、74（13.6%）、16（2.9%）和 14（2.6%）只蜱样本。检测伯氏疏螺旋体（Borrelia burgdorferi）最敏感的方法是针对 fla 基因的 PCR。