Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology
Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology
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DOI：
10.1128/jcm.35.4.907-914.1997
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发表时间：
1997-04-01
影响因子：
9.4
通讯作者：
vanEmbden, J
中科院分区：
文献类型：
--
作者：
Kamerbeek, J;Schouls, L;vanEmbden, J
DNA限制性片段长度多态性（RFLP）的广泛使用，以区分结核分枝杆菌菌株监测结核病的传播受到了阻碍，需要培养这种生长缓慢的有机体和RFLP分型所需的技术复杂程度。我们已经开发了一种简单的方法，可以同时检测和分型M。本发明的目的是在临床标本中检测结核病，并将怀疑疾病和分型之间的时间从1个月或几个月缩短到1或2天。该方法基于染色体DR基因座的多态性，其包含散布有非重复间隔区的可变数目的短同向重复序列。该方法被称为间隔区寡聚分型或“间隔区寡聚分型”，因为它基于具有多个间隔区寡核苷酸的体外扩增DNA的菌株依赖性杂交模式。大多数测试的临床分离株显示独特的杂交模式，而爆发菌株具有相同的spoligotype。直接检测临床标本所得的分型与用培养的M.结核这项新的初步研究表明，这种新的方法可能是一种有用的工具，用于快速披露社区、医院或其他机构中的相关暴发病例，并用于监测多重耐药支原体的传播。结核出乎意料的是，发现spoligotyping区分M。bovis从M.结核病，这是一个传统方法往往很难区分。
Widespread use of DNA restriction fragment length polymorphism (RFLP) to differentiate strains of Mycobacterium tuberculosis to monitor the transmission of tuberculosis has been hampered by the need to culture this slow-growing organism and by the level of technical sophistication needed for RFLP typing. We have developed a simple method which allows simultaneous detection and typing of M. tuberculosis in clinical specimens and reduces the time between suspicion of the disease and typing from 1 or several months to 1 or 2 days. The method is based on polymorphism of the chromosomal DR locus, which contains a variable number of short direct repeats interspersed with nonrepetitive spacers, The method is referred to as spacer oligotyping or ''spoligotyping'' because it is based on strain-dependent hybridization patterns of in vitro-amplified DNA with multiple spacer oligonucleotides, Most of the clinical isolates tested showed unique hybridization patterns, whereas outbreak strains shared the same spoligotype. The types obtained from direct examination of clinical samples were identical to those obtained by using DNA from cultured M. tuberculosis. This novel preliminary study shows that the novel method may be a useful tool for rapid disclosure of linked outbreak cases in a community, in hospitals, or in other institutions and for monitoring of transmission of multidrug-resistant M. tuberculosis. Unexpectedly, spoligotyping was found to differentiate M. bovis from M. tuberculosis, a distinction which is often difficult to make by traditional methods.