Clinical evaluation of mtp40 polymerase chain reaction for the diagnosis of extra pulmonary tuberculosis
- 1 December 2013
- journal article
- Published by Springer Science and Business Media LLC in World Journal of Microbiology and Biotechnology
- Vol. 30 (5), 1485-1490
- https://doi.org/10.1007/s11274-013-1566-z
Abstract
Rapid and sensitive detection of Mycobacterium tuberculosis from patient samples is vital for clinical diagnosis and treatment. The emergence of M. tuberculosis strains with either no copies or only a single copy of IS6110 in Asian countries makes the standard PCR based diagnosis of M. tuberculosis using IS6110 not reliable. We studied the diagnostic efficacy of the in-house PCR amplification of the candidate gene mtp40 as an alternative to IS6110 element based diagnosis. Clinical samples included pulmonary and extra-pulmonary specimens from TB suspected patients residing in Puducherry, South India and were analyzed using in-house PCR procedures targeting IS6110 element and mtp40 genes. Out of 317 clinical specimens analyzed, 132 (41.6 %) and 114 (36 %) were found positive for mtp40 PCR and IS6110 PCR, respectively. However, 18 specimens that were found to negative for IS6110 PCR were found positive for mtp40 PCR, which was further confirmed by DNA sequencing method. PCR amplification of mtp40 gene for the diagnosis of M. tuberculosis in clinical samples is fast, sensitive, and further identified clinical strains that lack IS6110 element in this region. It is clearly demonstrated that there is a significant difference between the two PCR procedures and the sensitivity and specificity levels of mtp40 PCR were found to be higher when compared with DNA sequencing method. Thus, mtp40 based PCR technique will be beneficial in diagnosis of TB where M. tuberculosis strains lack of IS6110 element is predominant.Keywords
This publication has 14 references indexed in Scilit:
- An in-house multiplex PCR test for the detection of Mycobacterium tuberculosis, its validation & comparison with a single target TB-PCR kit2012
- Molecular Epidemiological Study of Pyrazinamide-Resistance in Clinical Isolates of Mycobacterium tuberculosis from South IndiaInternational Journal of Molecular Sciences, 2010
- An Integrated Approach to Rapid Diagnosis of Tuberculosis and Multidrug Resistance Using Liquid Culture and Molecular Methods in RussiaPLOS ONE, 2009
- Diagnosis and Species Identification of Mycobacterial Infections by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis of Sterile Body FluidsThe Korean Journal of Internal Medicine, 2009
- Sensitivity of IS6110, mtp40 and 85B-RNA Based Amplification Assays in the Diagnosis and Treatment Follow up of Pulmonary Mycobacterium tuberculosisJournal of Biological Sciences, 2005
- Analysis of genetic polymorphisms affecting the four phospholipase C (plc) genes in Mycobacterium tuberculosis complex clinical isolatesMicrobiology, 2004
- Implications of Low Frequency of IS 6110 in Fingerprinting Field Isolates of Mycobacterium tuberculosis from Kerala, India Journal of Clinical Microbiology, 2001
- Clinical utility of the polymerase chain reaction in the diagnosis of extrapulmonary tuberculosisEuropean Respiratory Journal, 1998
- IS6110 restriction fragment length polymorphism typing of clinical isolates of Mycobacterium tuberculosis from patients with pulmonary tuberculosis in Madras, South IndiaTubercle and Lung Disease, 1995
- Evaluation of a polymerase chain reaction for the diagnosis of tuberculosisTubercle, 1991