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一种简单而高效的多重 PCR 检测方法，可用于鉴定分枝杆菌属和结核分枝杆菌复合体到种的水平。

A simple and efficient multiplex PCR assay for the identification of Mycobacterium genus and Mycobacterium tuberculosis complex to the species level.

机构信息

Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, 1 Yonseidae-gil, Wonju 220-710, Korea.

出版信息

Yonsei Med J. 2013 Sep;54(5):1220-6. doi: 10.3349/ymj.2013.54.5.1220.

DOI:10.3349/ymj.2013.54.5.1220
PMID:23918573
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3743195/
Abstract

PURPOSE

The Mycobacterium tuberculosis complex comprises M. tuberculosis, M. bovis, M. bovis bacillus Calmette-Guérin (BCG) and M. africanum, and causes tuberculosis in humans and animals. Identification of Mycobacterium spp. and M. tuberculosis complex to the species level is important for practical use in microbiological laboratories, in addition to optimal treatment and public health.

MATERIALS AND METHODS

A novel multiplex PCR assay targeting a conserved rpoB sequence in Mycobacteria spp., as well as regions of difference (RD) 1 and RD8, was developed and evaluated using 37 reference strains and 178 clinical isolates.

RESULTS

All mycobacterial strains produced a 518-bp product (rpoB), while other bacteria produced no product. Virulent M. tuberculosis complex strains, M. tuberculosis, M. bovis and M. africanum, produced a 254-bp product (RD1), while M. bovis BCG, M. microti and nontuberculous mycobacteria produced no RD1 region product. Additionally, M. tuberculosis and M. africanum produced a 150-bp product (RD8), while M. bovis and M. bovis BCG produced a 360-bp product (deleted form of RD8). M. microti and nontuberculous mycobacteria produced no RD8 region product. This assay identified all Mycobacterium spp. and all M. tuberculosis complex strains to the species level.

CONCLUSION

The multiplex PCR assay of the present study could be implemented as a routine test in microbiology laboratories, and may contribute to more effective treatment and surveillance of tuberculosis stemming from the M. tuberculosis complex.

摘要

目的

结核分枝杆菌复合体包括结核分枝杆菌、牛分枝杆菌、卡介苗（BCG）和非洲分枝杆菌，可引起人类和动物的结核病。除了优化治疗和公共卫生外，将分枝杆菌属和结核分枝杆菌复合体鉴定到种水平对于微生物实验室的实际应用也很重要。

材料和方法

本研究开发并评估了一种针对分枝杆菌属保守 rpoB 序列以及差异区（RD）1 和 RD8 的新型多重 PCR 检测方法，共使用 37 株参考菌株和 178 株临床分离株进行检测。

结果

所有分枝杆菌菌株均产生 518-bp 产物（rpoB），而其他细菌则未产生产物。毒力结核分枝杆菌复合体菌株结核分枝杆菌、牛分枝杆菌和非洲分枝杆菌产生 254-bp 产物（RD1），而卡介苗、鸟分枝杆菌和非结核分枝杆菌则未产生 RD1 区产物。此外，结核分枝杆菌和非洲分枝杆菌产生 150-bp 产物（RD8），而牛分枝杆菌和卡介苗则产生 360-bp 产物（RD8 的缺失形式）。鸟分枝杆菌和非结核分枝杆菌则未产生 RD8 区产物。本检测方法可将所有分枝杆菌属和所有结核分枝杆菌复合体菌株鉴定到种水平。

结论

本研究的多重 PCR 检测方法可作为微生物实验室的常规检测方法，有助于更有效地治疗和监测结核分枝杆菌复合体引起的结核病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/490fd273c940/ymj-54-1220-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/2921bcd0bb4d/ymj-54-1220-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/ce09afa43d86/ymj-54-1220-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/85bc2085f169/ymj-54-1220-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/1797ee8ac7e2/ymj-54-1220-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/490fd273c940/ymj-54-1220-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/2921bcd0bb4d/ymj-54-1220-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/ce09afa43d86/ymj-54-1220-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/85bc2085f169/ymj-54-1220-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/1797ee8ac7e2/ymj-54-1220-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4905/3743195/490fd273c940/ymj-54-1220-g005.jpg

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