Novel multiplex allele-specific PCR assays for the detection of resistance to second-line drugs in Mycobacterium tuberculosis
Novel multiplex allele-specific PCR assays for the detection of resistance to second-line drugs in Mycobacterium tuberculosis
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DOI：
10.1093/jac/dkq047
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发表时间：
2010-05-01
影响因子：
5.2
通讯作者：
Segal, Heidi
中科院分区：
文献类型：
--
作者：
Evans, Joanna;Segal, Heidi
使用快速分子检测法检测对二线药物的耐药性，将大大减少诊断耐药结核病（TB）的时间延迟，这与传统的表型药物敏感性测试有关。本研究采用多重等位基因特异性（MAS）PCR方法检测288株结核分枝杆菌GyrA D94 G和rrs A1401 G基因突变，并与氧氟沙星和卡那霉素进行比较。采用间接比例法对筛选出的耐多药结核菌株进行氧氟沙星和卡那霉素的表型药敏试验，MAS-PCR检测出表型耐药菌株中GyrA D94 G和rrs A1401 G突变，临床敏感性分别为54.5%（6/11）和90.0%（9/10），两种测定法的特异性均为100%。在11株氧氟沙星耐药分离株中有4株（36.4%）未携带D94 G置换，GyrA A90 V突变和rrs A1401 G突变的快速基因型检测可检出高达90.0%的西开普省广泛耐药（XDR）结核。在临床环境中使用这些检测方法将大大缩短诊断广泛耐药结核的时间，从而能够在治疗开始时实施适当的治疗方案。
The use of rapid molecular assays for the detection of resistance to second-line drugs would significantly decrease the time delay in diagnosing drug-resistant tuberculosis (TB) that is associated with conventional phenotypic drug susceptibility testing. In this study, multiplex allele-specific (MAS)-PCR assays designed to detect the GyrA D94G and rrs A1401G mutations were evaluated for detection of ofloxacin and kanamycin resistance.GyrA D94G and rrs A1401G MAS-PCR assays were carried out on 288 Mycobacterium tuberculosis isolates. Phenotypic drug susceptibility testing of ofloxacin and kanamycin was performed on selected multidrug-resistant TB isolates using the indirect proportions method.MAS-PCR assays detected GyrA D94G and rrs A1401G mutations in phenotypically resistant isolates with clinical sensitivities of 54.5% (6 of 11) and 90.0% (9 of 10), respectively, and specificities of 100% were obtained for both assays. A GyrA A90V mutation was identified in 4 of 11 (36.4%) ofloxacin-resistant isolates that did not carry a D94G substitution.Rapid genotypic assays designed to detect GyrA D94G and A90V mutations and rrs A1401G mutations could detect up to 90.0% of extensively drug-resistant (XDR)-TB in the Western Cape region. The use of these assays in the clinical setting would significantly reduce the time to diagnosis of XDR-TB, enabling the administration of appropriate treatment regimens at the outset of therapy.