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Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

机译:分枝杆菌迁移率变动分析:一种快速鉴定结核分枝杆菌和非结核分枝杆菌的方法

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The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.
机译:分枝杆菌的鉴定是必不可少的,因为结核病(TB)和分枝杆菌病在临床上无法区分,需要不同的治疗方案。传统的表型方法耗时,可能长达60天。确实,需要快速,负担得起的,特定的和易于执行的识别方法。我们之前已经描述了一种基于聚合酶链反应的方法,称为分枝杆菌迁移率变动分析(MMSA),该方法旨在用于结核分枝杆菌复合体(MTC)和非结核分枝杆菌(NTM)物种鉴定。这项研究的目的是评估MMSA,以鉴定MTC和NTM临床分离株,并将其性能与PRA-hsp65方法进行比较。 MMSA和PRA-hsp65共鉴定了204种临床分离株(102 NTM和102 MTC)。对于这些方法给出不一致结果的分离株,通过对16S rRNA和hsp65基因的片段进行测序,获得了明确的物种鉴定。两种方法都能正确识别所有MTC分离株。在NTM分离物中,单独的MMSA分配给一个复合物或物种94个(92.2%),而PRA-hsp65方法分配给一个物种100%。通过两种方法鉴定出的94种NTM分离株均观察到91.5%的一致性。 MMSA为96.8%的NTM分离物提供了正确的鉴定,而PRA-hsp65为94.7%。 MMSA是常规应用的合适辅助方法,用于快速鉴定分枝杆菌。

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