...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>The Indian journal of medical research. >Detection of Mycobacterium avium & M. tuberculosis from human sputum cultures by PCR-RFLP analysis of hsp65 gene & pncA PCR.
【24h】

Detection of Mycobacterium avium & M. tuberculosis from human sputum cultures by PCR-RFLP analysis of hsp65 gene & pncA PCR.

机译：通过hsp65基因的PCR-RFLP分析和pncA PCR检测人痰培养物中的鸟分枝杆菌和结核分枝杆菌。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

BACKGROUND & OBJECTIVE: Identification of mycobacteria by conventional methods is slow, labour intensive and may at times fail to produce precise results. Molecular techniques developed in the recent past, overcome these disadvantages facilitating rapid identification of most species. We undertook this study to characterize mycobacteria isolated from sputa of human patients suspected to have tuberculosis by conventional methods and later, by polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) of hsp65 gene and pncA PCR. METHODS: Twenty two mycobacteria isolated from 30 sputum samples were identified based on growth rate, pigmentation, cultural and biochemical properties and subjected to PRA of hsp65 gene involving amplification of hsp65 gene and digestion of the product with BstEII and HaeIII in separate reactions and analysis of digests by 3 per cent agarose gel electrophoresis. The mycobacteria were simultaneously evaluated by M. tuberculosis-specific and M. bovis-specific pncA PCR assays in separate reactions. RESULTS: With the conventional biochemical tests, the 22 sputum culture isolates were identified as M. tuberculosis (19) and M. avium complex (MAC) (3). PCR of hsp65 gene yielded 439 bp product in all the mycobacteria tested. The RFLP patterns of three MAC isolates with BstEII and HaeIII were identical to reference M. avium strain with two fragments in each of the digest. M. intracellulare reference strain showed a distinct pattern with 3 fragments each in both enzyme digests. The PRA of hsp65 confirmed MAC isolates as M. avium. M. tuberculosis isolates including H37Rv and M. bovis strains could not be discriminated by PRA of hsp65. The two pncA PCR assays (M. bovis-specific and M. tuberculosis-specific) detected specifically the respective organisms with an amplification product of 185 bp. The MAC strains yielded no amplification product in both the pncA PCR assays. INTERPRETATION & CONCLUSION: PRA profiles of hsp65 could differentiate MAC isolates into M. avium and M. intracellulare but could not distinguish between M. tuberculosis and M. bovis. pncA PCR assays were found specific in detecting the respective mycobacterial species. The study confirms utility of pncA PCR assays in differential identification of M. tuberculosis and M. bovis and that of PRA of hsp65 in the identification of M. avium.
机译：背景与目的：通过常规方法鉴定分枝杆菌的过程缓慢，劳动强度大，有时可能无法产生准确的结果。最近发展起来的分子技术克服了这些缺点，有助于快速鉴定大多数物种。我们进行了这项研究，以鉴定通过常规方法，然后通过hsp65基因的聚合酶链反应-限制性片段长度多态性分析（PRA）和pncA PCR从怀疑患有结核病的人类患者的痰中分离出的分枝杆菌。方法：根据生长速率，色素沉着，培养和生化特性鉴定了30份痰标本中的22株分枝杆菌，并对其进行了hsp65基因的PRA分析，涉及hsp65基因的扩增以及BstEII和HaeIII产物的消化，并分别进行了分析和分析。通过3％的琼脂糖凝胶电泳消化。分枝杆菌通过结核分枝杆菌特异性和牛分枝杆菌特异性pncA PCR分析同时在单独的反应中进行评估。结果：通过常规生化测试，鉴定出22种痰培养分离物为结核分枝杆菌（19）和鸟分枝杆菌复合体（MAC）（3）。在所有测试的分枝杆菌中，hsp65基因的PCR产生439 bp的产物。具有BstEII和HaeIII的三个MAC分离株的RFLP谱图与在每个消化物中均具有两个片段的参照鸟莫尔氏菌菌株相同。胞内分枝杆菌参考菌株显示出不同的模式，在两种酶消化物中每个都有3个片段。 hsp65的PRA证实MAC分离株为鸟分枝杆菌。 hsp65的PRA不能区分包括H37Rv和牛分枝杆菌在内的结核分枝杆菌。两种pncA PCR检测方法（牛分枝杆菌特异性和结核分枝杆菌特异性）分别检测到各自的微生物，其扩增产物为185 bp。在两种pncA PCR分析中，MAC菌株均未产生扩增产物。结论与结论：hsp65的PRA图谱可以将MAC分离株区分为鸟分枝杆菌和胞内分枝杆菌，但不能区分结核分枝杆菌和牛分枝杆菌。发现pncA PCR分析对检测各个分枝杆菌物种具有特异性。这项研究证实了pncA PCR分析在鉴别结核分枝杆菌和牛分枝杆菌以及hsp65的PRA在鉴别鸟分枝杆菌中的实用性。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号