@article{mbs:/content/journal/jmm/10.1099/jmm.0.000440, author = "Shrivastava, Kamal and Garima, Kushal and Narang, Anshika and Bhattacharyya, Kausik and Vishnoi, Ekta and Singh, Roshan Kumar and Chaudhry, Anil and Prasad, Rajendra and Bose, Mridula and Varma-Basil, Mandira", title = "Rv1458c: a new diagnostic marker for identification of Mycobacterium tuberculosis complex in a novel duplex PCR assay", journal= "Journal of Medical Microbiology", year = "2017", volume = "66", number = "3", pages = "371-376", doi = "https://doi.org/10.1099/jmm.0.000440", url = "https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.000440", publisher = "Microbiology Society", issn = "1473-5644", type = "Journal Article", keywords = "non-tuberculous mycobacteria", keywords = "duplex PCR assay", keywords = "Rv1458c", keywords = "rapid identification", keywords = "Mycobacterium tuberculosis complex", abstract = " Purpose. We explored the efficiency of Rv1458c, the gene encoding a putative ABC drug transporter specific for the Mycobacterium tuberculosis complex (MTBC), as a diagnostic marker. Methodology. A 190 bp region of Rv1458c and a 300 bp region of hsp65 were targeted in a novel duplex PCR assay and the results were compared with those for PCR restriction analysis(PRA) using the restriction enzymes NruI and BamHI. Species identification of a subset of the isolates (n=50) was confirmed by sequencing. Clinical isolates of M. tuberculosis (n=426) obtained from clinically suspected patients of pulmonary tuberculosis and mycobacterial (n=13) and non-mycobacterial (n=8) reference strains were included in the study. Results. The duplex PCR assay correctly identified 320/426 isolates as MTBC and 106/426 isolates as non-tuberculous mycobacteria(NTM). The test was 100 % specific and sensitive when compared with NruI/BamHI PCR restriction analysis and highlighted the use of Rv1458c as a diagnostic marker for MTBC. Conclusion. The duplex PCR assay could be developed for use as a screening test to identify MTBC in clinical specimens in peripheral laboratories with limited resources.", }