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首页> 外文期刊>Journal of Clinical Microbiology >Multiplex Real-Time PCR Assay for Rapid Identification of Mycobacterium tuberculosis Complex Members to the Species Level
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Multiplex Real-Time PCR Assay for Rapid Identification of Mycobacterium tuberculosis Complex Members to the Species Level

机译：多重实时荧光定量PCR快速鉴定结核分枝杆菌复杂成员的物种水平

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The species identification of members of the Mycobacterium tuberculosis complex is critical to the timely initiation of both appropriate antibiotic therapy and proper public health control measures. However, the current commercially available molecular assays identify mycobacteria only to the complex level and are unable to differentiate M. tuberculosis from the closely related M. bovis and M. bovis BCG. We describe here a rapid and robust two-step, multiplex, real-time PCR assay based on genomic deletions to definitively identify M. tuberculosis, M. bovis, M. bovis BCG, and other members of the complex. When tested against a panel of well-characterized mycobacterial reference strains, the assay was both sensitive and specific, correctly identifying all strains. We applied this assay to 60 clinical isolates previously identified as M. tuberculosis complex and found 57 M. tuberculosis isolates and 3 M. bovis BCG isolates from patients who had received intravesical BCG. Furthermore, analysis of 15 clinical specimens previously identified as M. bovis by spoligotyping revealed an isolate of M. tuberculosis that had been misidentified. We propose that this assay will allow the routine identification of M. tuberculosis complex members in the clinical laboratory.
机译：结核分枝杆菌复合物成员的物种识别对于及时启动适当的抗生素治疗和适当的公共卫生控制措施至关重要。但是，当前可商购的分子测定法只能在复杂水平上鉴定分枝杆菌，不能区分 M。与结核病密切相关的 M bovis 和 M。 bovis BCG。我们在这里描述了一种基于基因组缺失的快速而稳健的两步，多重，实时PCR分析法，以明确鉴定 M。结核菌，牛分枝杆菌，牛分枝杆菌 BCG和其他复合物。当针对一组特征明确的分枝杆菌参考菌株进行测试时，该测定既灵敏又特异性，可以正确鉴定所有菌株。我们将此分析应用于60个先前鉴定为 M的临床分离株。结核复合物，发现57个 M。结核病分离物和3个 M。 bovis 卡介苗分离自接受膀胱内BCG的患者。此外，分析了之前鉴定为 M的15个临床标本。 Bovis 通过spoligotyping发现了 M的分离株。结核病被误认了。我们建议该测定法将允许常规鉴定 M。结核复杂的成员在临床实验室。

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