Journal Article

Evaluation of a Polymerase Chain Reaction-Based Universal Heteroduplex Generator Assay for Direct Detection of Rifampin Susceptibility of <i>Mycobacterium tuberculosis</i> from Sputum Specimens

Diana L. Williams, Laynette Spring, Thomas P. Gillis, Max Salfinger and David H. Persing

in Clinical Infectious Diseases

Published on behalf of Infectious Diseases Society of America

Volume 26, issue 2, pages 446-450
Published in print February 1998 | ISSN: 1058-4838
Published online February 1998 | e-ISSN: 1537-6591 | DOI: http://dx.doi.org/10.1086/516313
Evaluation of a Polymerase Chain Reaction-Based Universal Heteroduplex Generator Assay for Direct Detection of Rifampin Susceptibility of Mycobacterium tuberculosis from Sputum Specimens

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In a double-blind study, 655 sputum specimens were obtained from individuals suspected of having tuberculosis and were analyzed for the presence of Mycobacterium tuberculosis and rifampin susceptibility with use of a polymerase chain reaction (PCR)-based universal heteroduplex generator assay (PCR/UHG-Rif). Of the specimens containing viable M. tuberculosis, 100% of the smearpositive (nn = 41) and 50% of the smear-negative (n = 6) specimens tested positive for the organism by PCR/UHG-Rif. Nineteen of 537 culture-negative specimens tested positive for M. tuberculosis by PCR/UHG-Rif and were from patients with confirmed tuberculosis who were receiving antituberculosis therapy at the time of specimen collection. Thirty-five specimens contained nontuberculous mycobacteria and were negative by PCR/UHG-Rif. Genotypic evidence of rifampin resistance in five of six culture-confirmed, rifampin-resistant isolates was obtained by PCR/UHG-Rif, yielding a sensitivity and specificity for the assay of 83% and 98.2%, respectively. These results demonstrate the feasibility of using a PCR-based assay directly on sputum specimens for simultaneous detection of M. tuberculosis and rifampin susceptibility, and they suggest that patients with smear-positive, untreated tuberculosis and those presenting with suspected drug-resistant tuberculosis are the most appropriate groups for testing by PCR/UHG-Rif.

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Subjects: Infectious Diseases ; Immunology ; Public Health and Epidemiology ; Microbiology

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