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应用罕见限制位点扩增技术对结核分枝杆菌和非结核分枝杆菌进行基因分型。

Application of infrequent-restriction-site amplification for genotyping of Mycobacterium tuberculosis and non-tuberculous mycobacterium.

作者信息

Choi Tae Yeal, Kang Jung Oak

机构信息

Department of Clincal Pathology, Hanyang University Medical College, Sung-dung-gu, Seoul, Korea.

出版信息

J Korean Med Sci. 2002 Oct;17(5):593-8. doi: 10.3346/jkms.2002.17.5.593.

DOI:10.3346/jkms.2002.17.5.593
PMID:12378007
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3054925/
Abstract

Infrequent restriction site amplification (IRS-PCR) is a method of amplifying DNA sequences, which flank an infrequent restriction site, and produces a strain-specific electrophoretic pattern. We studied the use of IRS-PCR to characterize Mycobacterium tuberculosis and non-tuberculous mycobactria (NTM). One-hundred and sixteen M. tuberculosis and nine NTM isolated at Hanyang University Hospital in Seoul, Korea were used in this study. IRS-PCR using AH1 and PX-G primers produced unique patterns for reference strains, M. tuberculosis H37Rv, M. bovis BCG, M. kansasii, M. scrofulaceum, M. szulgai, M. gordonae, M. avium, M. intracellulae, M. fortuitum, and M. chelonae, respectively. Reference strains M. tuberculosis H37Rv, M. bovis, M. africanum, and all isolates of M. tuberculosis showed similar IRS-PCR patterns. The IRS-PCR patterns generated with multiple isolates of M. tuberculosis from the same patients were essentially identical. IRS-PCR revealed the greatest difference between electrophoretic DNA patterns from M. avium, M. intracellulae, and M. fortuitum that differed from each other and from the reference strains. We concluded that IRS-PCR is a useful tool for strain typing of NTM, but not for M. tuberculosis.

摘要

稀有酶切位点扩增（IRS-PCR）是一种扩增位于稀有酶切位点侧翼的DNA序列的方法，可产生菌株特异性的电泳图谱。我们研究了IRS-PCR在结核分枝杆菌和非结核分枝杆菌（NTM）鉴定中的应用。本研究使用了韩国首尔汉阳大学医院分离的116株结核分枝杆菌和9株NTM。使用AH1和PX-G引物的IRS-PCR分别为参考菌株结核分枝杆菌H37Rv、牛分枝杆菌卡介苗、堪萨斯分枝杆菌、瘰疬分枝杆菌、苏尔加分枝杆菌、戈登分枝杆菌、鸟分枝杆菌、胞内分枝杆菌、偶然分枝杆菌和龟分枝杆菌产生了独特的图谱。参考菌株结核分枝杆菌H37Rv、牛分枝杆菌、非洲分枝杆菌以及所有结核分枝杆菌分离株均显示出相似的IRS-PCR图谱。来自同一患者的多个结核分枝杆菌分离株产生的IRS-PCR图谱基本相同。IRS-PCR显示，鸟分枝杆菌、胞内分枝杆菌和偶然分枝杆菌之间以及与参考菌株之间的电泳DNA图谱差异最大。我们得出结论，IRS-PCR是NTM菌株分型的有用工具，但不适用于结核分枝杆菌。