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首页> 外文期刊>Journal of Clinical Microbiology >Heterogeneity of RNA Polymerase Gene (rpoB) Sequences of Mycobacterium gordonae Clinical Isolates Identified with a DNA Probe Kit and by Conventional Methods
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Heterogeneity of RNA Polymerase Gene (rpoB) Sequences of Mycobacterium gordonae Clinical Isolates Identified with a DNA Probe Kit and by Conventional Methods

机译：用DNA探针试剂盒和常规方法鉴定的戈登分枝杆菌临床分离株RNA聚合酶基因（rpoB）序列的异质性

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In a previous study, we have evaluated genetic identification by using the rpoB gene, which was recently introduced by Kim et al. (J. Clin. Microbiol. 39:2102-2109, 2001; J. Clin. Microbiol. 37:1714-1720, 1999). In this process, we examined the rpoB gene heterogeneity of clinical isolates identified as Mycobacterium gordonae with the conventional biological and biochemical tests and/or a commercially available DNA probe kit. Sequencing of the rpoB gene of 34 clinical isolates revealed that M. gordonae clinical isolates were classified into four major clusters (A, B, C, and D). Interestingly, organisms belonging to cluster D (15 isolates) did not hybridize with M. gordonae ATCC 14470 and specifically possessed urease activity. Therefore, it could be considered to be a novel mycobacterium. The identification of M. gordonae is known to have ambiguous results sometimes. On the other hand, identification of clinical isolates seems to be inconvenient and unsuitable because of a more than 99% 16S rRNA gene similarity value between clusters. These findings suggest that the existence of M. gordonae-like mycobacteria that share similar biochemical and biological characteristics with the 16S rRNA gene of an M. gordonae type strain but less similarity at the genomic DNA level may have complicated the identification of M. gordonae in many laboratories. Furthermore, compared with hsp65 PCR restriction analysis (PRA), rpoB PRA would have the advantage of producing no ambiguous results because of the intracluster homogeneity of the rpoB gene. In this case, rpoB would provide clearer results than hsp65, even if PRA analysis was used. We demonstrated that these M. gordonae-like mycobacteria were easily distinguished by PRA of the rpoB sequence. Additionally, the significance of this M. gordonae-like cluster may help to establish the comparison between the M. gordonae isolates from a clinical specimen and an infectious process in a given patient and to determine the true incidence of infection with this microorganism.
机译：在先前的研究中，我们使用Kim等人最近引入的 rpoB 基因评估了遗传鉴定。 （J.Clin.Microbiol.39：2102-2109，2001; J.Clin.Microbiol.37：1714-1720，1999）。在此过程中，我们使用常规的生物学和生化测试和/或可商购的DNA探针试剂盒检查了被鉴定为戈登分枝杆菌的临床分离株的rpoB基因异质性。对34个临床分离株的 rpoB 基因进行测序后发现， M。 gordonae 临床分离株分为四个主要簇（A，B，C和D）。有趣的是，属于簇D的生物（15个分离株）未与 M杂交。 gordonae ATCC 14470，特别具有脲酶活性。因此，可以认为它是新型分枝杆菌。 M的标识。众所周知，gordonae 具有不明确的结果。另一方面，由于分离株之间的16S rRNA基因相似性值超过99％，因此临床分离株的鉴定似乎不方便且不合适。这些发现表明 M的存在。类似于戈登菌的分枝杆菌，具有与 M的16S rRNA基因相似的生化和生物学特性。 gordonae 型菌株，但在基因组DNA水平上的相似性较低，可能使 M的鉴定变得复杂。实验室中的毛don科。此外，与 hsp65 PCR限制分析（PRA）相比， rpoB PRA的优势在于不会产生歧义的结果，因为 rpoB 的簇内均一性基因。在这种情况下，即使使用PRA分析， rpoB 也将提供比 hsp65 更清晰的结果。我们证明了这些 M。通过 rpoB 序列的PRA可以很容易地区分出戈登菌样分枝杆菌。此外，此 M的意义。类gordonae 簇可能有助于建立 M之间的比较。戈登菌从特定患者的临床标本和感染过程中分离出来，并确定该微生物感染的真实发生率。

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