A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries
A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries
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DOI：
10.1099/jmm.0.2008/002600-0
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发表时间：
2008-10-01
影响因子：
3
通讯作者：
Suzuki, Yasuhiko
中科院分区：
文献类型：
--
作者：
Matsuoka, Masanori;Aye, Khin Saw;Suzuki, Yasuhiko
利用反向杂交系统，建立了一种简单的方法来检测麻风分枝杆菌基因组中对麻风关键药物产生耐药性的突变。选择与麻风对氨苯砜、利福平和氧氟沙星耐药相关的folP 1、rpoB和gyrA基因突变相对应的寡核苷酸探针，固定在载玻片上作为捕获探针，构建了麻风药物敏感性-DNA微阵列（LDS-DA）。临床分离的M.在缅甸和菲律宾这两个发展中国家进行了可行性研究，以评估最不发达国家发展援助方案的绩效。通过该方法获得的结果与核苷酸测序结果的高度一致性强烈支持LDS-DA作为药物敏感性测试代替测序的适用性，测序是一种耗时且昂贵的程序。该方法快速、简便，可同时进行三种一线药物的敏感性检测，解决了以往方法存在的问题。
A simple method to detect mutations in the genome of Mycobacterium leprae that confer resistance to key drugs for leprosy was exploited on the basis of a reverse hybridization system. A series of oligonucleotide probes corresponding to each mutation in the folP1, rpoB and gyrA genes for dapsone, rifampicin and ofloxacin resistance, respectively, were selected and fixed on a glass slide as capture probes, to develop a DNA microarray termed the leprosy drug susceptibility-DNA microarray (LDS-DA). Mutations in clinical isolates of M. leprae were successfully identified by the LDS-DA. Feasibility studies were conducted to evaluate the performance of the LDS-DA in two developing countries, Myanmar and the Philippines. The high concordance of the results obtained by this method with the results of nucleotide sequencing strongly supports the applicability of the LDS-DA as a drug susceptibility test in place of sequencing, a time-consuming and costly procedure. This is a rapid and simple method for the simultaneous susceptibility testing of three front-line drugs for leprosy, and solves the problems of previously reported methods.