Pyrosequencing identification of Mycobacterium tuberculosis W-Beijing.
Pyrosequencing identification of Mycobacterium tuberculosis W-Beijing.
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DOI：
10.1186/1756-0500-2-239
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发表时间：
2009-12-02
影响因子：
1.8
通讯作者：
Drancourt M
中科院分区：
文献类型：
--
作者：
Djelouadji Z;Henry M;Bachtarzi A;Foselle N;Raoult D;Drancourt M
全球范围内不断扩大的结核分枝杆菌W-Beijing家族与治疗失败和复发有关。目前，该病毒的鉴定依赖于基因分型和常规测序。我们开发了焦磷酸测序作为鉴定它的另一种方法。焦磷酸测序发现Rv0927c-pstS3基因间隔区G/A替换和RD105缺失，鉴定出8/104株结核分枝杆菌为W-Beijing分离株。此外，焦磷酸测序发现W-Beijing分离株的Rv0927c基因存在以前未报道的TGC缺失。焦磷酸测序数据与常规测序数据以及参考分子鉴定结果完全一致。多间隔序列分析将W-Beijing分离株归入亚洲血统，将96株非W-Beijing分离株归类为欧美血统(P<；10-5)。W-Beijing分离株对链霉素、利福平、异烟肼、乙胺丁醇和吡嗪酰胺均敏感，未检测到耐药相关突变。W-Beijing与非W-Beijing分离株对抗生素的敏感性差异无统计学意义(p=0.2，X2检验)。焦磷酸测序法在26例显示抗酸杆菌的痰标本中正确地鉴定出结核分枝杆菌。焦磷酸测序结果是在四个小时内获得的，估计每项测试的成本为1.86欧元。对Rv0927c基因和相邻的基因间隔区进行焦磷酸测序是一种高效、低成本的快速鉴定W-Beijing分离株的技术。
The worldwide expanding Mycobacterium tuberculosis W-Beijing family is associated with treatment failure and relapse. Its identification currently relies on spoligotyping and conventional sequencing. We developed pyrosequencing as an alternative method for its identification. Pyrosequencing found a G/A substitution in the Rv0927c-pstS3 intergenic spacer and a RD105 deletion, identifying 8/104 M. tuberculosis isolates as W-Beijing isolates. In addition, pyrosequencing found a previously unreported TGC deletion in the Rv0927c gene of W-Beijing isolates. Total concordance was found between the pyrosequencing data and conventional sequencing, as well as reference molecular identification. Multispacer Sequence Typing assigned the W-Beijing isolates to the Asian lineage and the 96 non-W-Beijing isolates to the Euro-American lineage (P < 10-5). The W-Beijing isolates were all susceptible to streptomycin, rifampin, isoniazid, ethambutol, and pyrazinamide; no resistance-associated mutations were detected in these eight W-Beijing isolates. There were no statistically significant differences in the antibiotic susceptibility of W-Beijing and non-W-Beijing isolates (p = 0.2, X2 test). Pyrosequencing correctly identified M. tuberculosis organisms in 26/26 sputum specimens exhibiting acid-fast bacilli. Pyrosequencing results were obtained within four hours, incurring an estimated cost of 1.86 €/test. Pyrosequencing of the Rv0927c gene and adjacent intergenic spacer is an efficient, low-cost technique for the rapid identification of W-Beijing isolates.