A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP
A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP
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DOI：
10.1016/j.micinf.2009.01.008
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发表时间：
2009-03-01
影响因子：
5.8
通讯作者：
Zhang, Ying
中科院分区：
文献类型：
--
作者：
Jiang, Xin;Lu, Chanyi;Zhang, Ying
最近我们发现W-北京结核分枝杆菌菌株在Rv 0927 c的421位有一个独特的读码框内三核苷酸（in-frame trinucleotide，AGC）缺失，在Rv 0927 c-pstS 3基因间区有一个-127G → A突变。在检测到这两个突变的421个三核苷酸缺失的基础上，我们建立了一种快速鉴定非北京菌株中的W-北京菌株的PCR-SSCP方法。总共分析了104株临床分离株，其中包括68株西北京菌株和36株非北京菌株。我们发现PCR-SSCP成功地区分了所有的W-北京菌株和非北京菌株。另外，我们意外地发现SDS-PAGE蛋白质凝胶在检测AGC缺失突变的SSCP分析中具有比传统的TBE聚丙烯酰胺凝胶更好的分辨率。(C)2009年由Elsevier Masson SAS出版。
Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a -127G -> A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide get in detecting the AGC deletion mutation in the SSCP analysis. (C) 2009 Published by Elsevier Masson SAS.