Identification of mycobacterial species by PCR restriction enzyme analysis of the hsp65 gene—an Indian experience.
Identification of mycobacterial species by PCR restriction enzyme analysis of the hsp65 gene—an Indian experience.
复制标题
通过对 hsp65 基因进行 PCR 限制性内切酶分析来鉴定分枝杆菌物种——印度的经验。
DOI：
10.1139/cjm-2014-0525
复制
发表时间：
2015
影响因子：
2.8
通讯作者：
Sarin,Rohit
中科院分区：
文献类型：
--
作者：
Verma,AjoyKumar;Kumar,Gavish;Arora,Jyoti;Singh,Paras;Arora,VijayKumar;Myneedu,VithalPrasad;Sarin,Rohit
目前，非结核分枝杆菌(NTM)常引起肺和肺外疾病。NTM的种类鉴定决定了该病的治疗和管理路线。非结核分枝杆菌的常规诊断方法，即涂片显微镜和生化鉴定，既繁琐又耗时，并不是所有的实验室都能常规进行这些测试。用标准菌株进行针对hsp65基因的聚合酶链式反应，并对109株临床分离株进行了检测。扩增产物用BstEII和HaeIII进行限制性内切酶分析。并将所得结果与生化试验结果进行比较。在109个NTM中，有107个被鉴定为物种水平。聚合酶链式反应加限制性内切酶分析鉴定出12种NTM类型。在生长缓慢的非结核分枝杆菌中，常见的种类有龟分枝杆菌(32株)和禽型分枝杆菌复合体(21株)。PRA与生化鉴定的符合率为95.32%(102/107)。PRA是一种快速、廉价、准确的诊断潜在致病性NTM的方法。
Nowadays, nontuberculous mycobacteria (NTM) often cause pulmonary and extrapulmonary disease. Species identification of NTM determines the line of treatment and management of the disease. The routine diagnostic methods, i.e., smear microscopy and biochemical identification, of nontuberculous mycobacteria are tedious and time consuming and not all laboratories can perform these tests on a routine basis. A PCR targeting thehsp65gene was implemented using standard strains and was applied to 109 clinical isolates. The PCR-amplified product was subjected to restriction enzyme analysis usingBstEII andHaeIII. The results obtained were compared with that of biochemical tests. Of 109 NTM, 107 were identified to species level. PCR plus restriction enzyme analysis (PRA) identified 12 types of NTM. Common species identified wereMycobacterium chelonae(32), a rapid growing NTM, andMycobacterium aviumcomplex (21), among the slow growing NTM. PRA and biochemical identification showed 95.32% (102/107) concordant results. PRA is fast, cheap, and accurate for identification of potentially pathogenic NTM.