Kahla Imen Ben, Henry Mireille, Boukadida Jalel, Drancourt Michel
URMITE, CNRS UMR6236, IRD198, IFR 48, Institut Méditerranée Infection, Aix-Marseille-Université, Marseille, France.
BMC Res Notes. 2011 Oct 19;4:423. doi: 10.1186/1756-0500-4-423.
Identification of the Mycobacterium tuberculosis complex organisms to the species level is important for diagnostic, therapeutic and epidemiologic perspectives. Indeed, isolates are routinely identified as belonging to the M. tuberculosis complex without further discrimination in agreement with the high genomic similarity of the M. tuberculosis complex members and the resulting complex available identification tools.
We herein develop a pyrosequencing assay analyzing polymorphisms within glpK, pykA and gyrB genes to identify members of the M. tuberculosis complex at the species level. The assay was evaluated with 22 M. tuberculosis, 21 M. bovis, 3 M. caprae, 3 M. microti, 2 M. bovis BCG, 2 M. pinnipedii, 1 M. canettii and 1 M. africanum type I isolates. The resulted pyrograms were consistent with conventional DNA sequencing data and successfully identified all isolates. Additionally, 127 clinical M. tuberculosis complex isolates were analyzed and were unambiguously identified as M. tuberculosis.
We proposed a pyrosequencing-based scheme for the rapid identification of M. tuberculosis complex isolates at the species level. The assay is robust, specific, rapid and can be easily introduced in the routine activity.
从诊断、治疗和流行病学角度来看，将结核分枝杆菌复合群微生物鉴定到种水平很重要。实际上，由于结核分枝杆菌复合群成员的高基因组相似性以及现有的复合群鉴定工具，分离株通常被常规鉴定为属于结核分枝杆菌复合群，而无需进一步区分。
我们在此开发了一种焦磷酸测序分析方法，用于分析glpK、pykA和gyrB基因内的多态性，以在种水平上鉴定结核分枝杆菌复合群的成员。该分析方法用22株结核分枝杆菌、21株牛分枝杆菌、3株山羊分枝杆菌、3株田鼠分枝杆菌、2株卡介苗、2株海兽分枝杆菌、1株堪萨斯分枝杆菌和1株Ⅰ型非洲分枝杆菌分离株进行了评估。所得的焦磷酸测序图与传统DNA测序数据一致，并成功鉴定了所有分离株。此外，对127株临床结核分枝杆菌复合群分离株进行了分析，并明确鉴定为结核分枝杆菌。
我们提出了一种基于焦磷酸测序的方案，用于在种水平上快速鉴定结核分枝杆菌复合群分离株。该分析方法稳健、特异、快速，并且可以很容易地引入常规检测中。
