Mistri S K, Sultana M, Kamal S M M, Alam M M, Irin F, Nessa J, Ahsan C R, Yasmin M
Department of Microbiology, University of Dhaka, Dhaka, Bangladesh.
National Tuberculosis Reference Laboratory, NIDCH, Dhaka, Bangladesh.
Lett Appl Microbiol. 2016 May;62(5):411-8. doi: 10.1111/lam.12564.
For an effective control of tuberculosis, rapid detection of multidrug resistant tuberculosis (MDR-TB) is necessary. Therefore, we developed a modified nested multiplex allele-specific polymerase chain reaction (MAS-PCR) method that enables rapid MDR-TB detection directly from sputum samples. The efficacy of this method was evaluated using 79 sputum samples collected from suspected tuberculosis patients. The performance of nested MAS-PCR method was compared with other MDR-TB detection methods like drug susceptibility testing (DST) and DNA sequencing. As rifampicin (RIF) resistance conforms to MDR-TB in greater than 90% cases, only the presence of RIF-associated mutations in rpoB gene was determined by DNA sequencing and nested MAS-PCR to detect MDR-TB. The concordance between nested MAS-PCR and DNA sequencing results was found to be 96·3%. When compared with DST, the sensitivity and specificity of nested MAS-PCR for RIF-resistance detection were determined to be 92·9 and 100% respectively.
For developing- and high-TB burden countries, molecular-based tests have been recommended by the World Health Organization for rapid detection of MDR-TB. The results of this study indicate that, nested MAS-PCR assay might be a practical and relatively cost effective molecular method for rapid detection of MDR-TB from suspected sputum samples in developing countries with resource poor settings.
为有效控制结核病，快速检测耐多药结核病（MDR-TB）很有必要。因此，我们开发了一种改良的巢式多重等位基因特异性聚合酶链反应（MAS-PCR）方法，可直接从痰标本中快速检测MDR-TB。使用从疑似结核病患者收集的79份痰标本评估了该方法的有效性。将巢式MAS-PCR方法的性能与其他MDR-TB检测方法如药物敏感性试验（DST）和DNA测序进行了比较。由于超过90%的病例中利福平（RIF）耐药符合MDR-TB，通过DNA测序和巢式MAS-PCR仅测定rpoB基因中与RIF相关的突变以检测MDR-TB。发现巢式MAS-PCR与DNA测序结果之间的一致性为96.3%。与DST相比，巢式MAS-PCR检测RIF耐药的敏感性和特异性分别确定为92.9%和100%。
对于结核病高负担国家和发展中国家，世界卫生组织推荐基于分子的检测方法用于快速检测MDR-TB。本研究结果表明，巢式MAS-PCR检测可能是一种实用且相对经济有效的分子方法，用于在资源匮乏的发展中国家从疑似痰标本中快速检测MDR-TB。
