Characterization of rpoB mutations in rifampin-resistant clinical Mycobacterium tuberculosis isolates from Greece
Characterization of rpoB mutations in rifampin-resistant clinical Mycobacterium tuberculosis isolates from Greece
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DOI：
10.1128/jcm.36.1.20-23.1998
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发表时间：
1998-01-01
影响因子：
9.4
通讯作者：
Marinis, E
中科院分区：
文献类型：
--
作者：
Matsiota-Bernard, P;Vrioni, G;Marinis, E
我们研究了17株来自希腊患者的利福平耐药临床分离株，以鉴定rpoB突变。我们研究的目的是评估市售的线探针检测试剂盒（ino - lipa Rif， TB）检测rpoB突变和利福平耐药性。将市售检测结果与PCR扩增产物的自动DNA序列分析结果进行了比较，并对分离株进行了随机扩增多态性DNA （RAPD）分析，线探针检测与表型利福平敏感性试验的总体一致性为94%，正确识别出密码子Ser（531）、His（526）和Asp（516）三个不同的rpoB突变。但是外部区域和插入的突变只能通过自动DNA序列分析检测到，密码子Ser（531）和His（526）的变化是利福平耐药的主要原因，正如之前对其他地理区域的分离株所描述的那样。对分离株进行RAPD分析的结果表明，克隆相关的结核分枝杆菌菌株可能具有携带不同突变rpoB等位基因的亚克隆。该方法快速且易于进行测试，可用于在常规方法获得结果之前快速检测结核分枝杆菌对利福平的耐药性，并用于流行病学研究，但该方法获得的阴性结果并不排除对利福平的耐药性。
There is a geographic distribution of Mycobacterium tuberculosis strains with various rpoB gene mutations that account for rifampin resistance, We studied 17 rifampin-resistant clinical isolates from patients in Greece to identify rpoB mutations, The aim of our study was the evaluation of a commercially available line probe assay kit (INNO-LiPA Rif, TB) to detect rpoB mutations and rifampin resistance, The results obtained with the commercially available assay were compared to those obtained by automated DNA sequence analysis of amplified PCR products, Randomly amplified polymorphic DNA (RAPD) analyses of the isolates were also performed, The overall concordance of the line probe assay with phenotypic rifampin susceptibility test was 94%, Three distinct rpoB mutations in codons Ser(531), His(526), and Asp(516) were correctly identified with the kit, but mutations in external regions and insertions were detected only by automated DNA sequence analysis, The changes in codons Ser(531) and His(526) accounted for the majority of rifampin resistance, as previously described for isolates from other geographic areas, The results obtained by RAPD analyses of the isolates suggested that clonally related M. tuberculosis strains can have subclones bearing distinct mutant rpoB alleles, We conclude that this line probe assay kit, which is fast and with which tests are easy to perform, can be used for the rapid detection of rifampin resistance in M, tuberculosis before the availability of results by conventional methods and for epidemiological studies but that negative results obtained by this method do not rule out rifampin resistance.