Standardization and evaluation of a tetraplex polymerase chain reaction to detect and differentiate Mycobacterium tuberculosis complex and nontuberculous Mycobacteria--a retrospective study on pulmonary TB patients |
| |
Authors: | Anilkumar A K Madhavilatha G K Paul Laiza K Radhakrishnan Indulakshmi Kumar R Ajay Mundayoor Sathish |
| |
Affiliation: | Mycobacterium Research Group, Department of Molecular Microbiology, Rajiv Gandhi Centre for Biotechnology, Trivandrum 695014, India. |
| |
Abstract: | The clinical presentation of pulmonary tuberculosis by members of Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) cannot be differentiated using the available standard diagnostic procedures. A single-tube tetraplex polymerase chain reaction (T-PCR) was designed to simultaneously amplify 4 well-known DNA targets of MTC. Taguchi's protocol was followed for the optimization of the conditions and was then tested on 288 pulmonary TB patient samples. The analytical sensitivity of the T-PCR was 100 fg of purified mycobacterial DNA, and specificity was found to be 100% in being able to distinguish MTC and NTM in all the cases tested. The results correlated well when validated with hsp65 PCR restriction analysis and sequencing of the 16S-23S internal transcribed spacer region, hsp65, and rpoB. The T-PCR described here is a quick, valuable, and cost-effective tool for determining whether the causative organism is MTC or NTM, and thus is useful for disease surveillance. |
| |
Keywords: | Mycobacterium tuberculosis complex NTM Multiplex PCR Taguchi's protocol Atypical Mycobacteria Disease surveillance |
本文献已被 ScienceDirect PubMed 等数据库收录! |