Biochemistry Research Laboratory, Central India Institute of Medical Sciences, 88/2, Bajaj Nagar, Nagpur, 440010 Maharashtra India.
Environmental Genomics Unit, National Environmental Engineering Research Institute, Nehru Marg, Nagpur, 440020 India.
Indian J Microbiol. 2015 Jun;55(2):213-8. doi: 10.1007/s12088-015-0517-9. Epub 2015 Feb 3.
The present study describes the development and evaluation of a duplex polymerase chain reaction (D-PCR) for diagnosis and simultaneous identification of tuberculous meningitis (TBM) and bacterial meningitis (BM) in a single reaction. A D-PCR with primers amplifying portions of the Mycobacterium tuberculosis IS6110 and the eubacteria 16SrDNA sequence in a same reaction mix was developed and tested on DNA extracted from 150 clinical CSF samples from different categories (TBM = 39, BM = 26, control infectious and non-infectious category = 85). The results indicate a clear differentiation between bands for eubacteria and M. tuberculosis with an analytical sensitivity of 10(3) cfu/ml for eubacteria and 10(2) cfu/ml for M. tuberculosis. When evaluated in clinical samples, D-PCR overall diagnosed 100 % confirmed TBM and 100 % confirmed BM cases with overall specificity of 96.5 %. D-PCR can be an effective tool for diagnosis and simultaneous identification of TBM or BM in a single PCR reaction. It saves time, cost, labour and sample amount and help in administration of appropriate antimicrobial therapy. The proposed diagnostic assay would be helpful in correct and rapid management of TBM and BM patients.
本研究描述了一种双重聚合酶链反应（D-PCR）的开发和评估，用于在单个反应中诊断结核性脑膜炎（TBM）和细菌性脑膜炎（BM）并进行同时鉴定。开发了一种 D-PCR，其引物在同一反应混合物中扩增分枝杆菌结核分枝杆菌 IS6110 和真核生物 16SrDNA 序列的部分，并在来自不同类别（TBM=39、BM=26、感染和非感染对照类别=85）的 150 个临床 CSF 样本的 DNA 上进行了测试。结果表明，真核生物和分枝杆菌之间的条带明显分化，真核生物的分析灵敏度为 10(3)cfu/ml，分枝杆菌的分析灵敏度为 10(2)cfu/ml。在临床样本中评估时，D-PCR 总体上诊断出 100%确诊的 TBM 和 100%确诊的 BM 病例，总体特异性为 96.5%。D-PCR 可以成为一种在单个 PCR 反应中诊断和同时鉴定 TBM 或 BM 的有效工具。它节省了时间、成本、劳动力和样本量，并有助于实施适当的抗菌治疗。该诊断检测方法将有助于正确和快速管理 TBM 和 BM 患者。
