title: Evaluation of a polymerase chain reaction-based universal heteroduplex generator assay for direct detection of rifampin susceptibility of Mycobacterium tuberculosis from sputum specimens.
abstract: 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 smear-positive (n = 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.
authors.affiliation: Molecular Biology Research Department, Gillis W. Long Hansen's Disease Center at Louisiana State University, Baton Rouge 70894, USA.
authors.forename: D L
authors.initials: DL
authors.lastname: Williams
authors.forename: L
authors.initials: L
authors.lastname: Spring
authors.forename: T P
authors.initials: TP
authors.lastname: Gillis
authors.forename: M
authors.initials: M
authors.lastname: Salfinger
authors.forename: D H
authors.initials: DH
authors.lastname: Persing
mesh_terms: D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D004269:DNA, Bacterial; D004311:Double-Blind Method; D004352:Drug Resistance, Microbial; D005069:Evaluation Studies as Topic; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D009692:Nucleic Acid Heteroduplexes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology*; D013183:Sputum / Q000382:microbiology
chemical_list: D000904:Antibiotics, Antitubercular; D004269:DNA, Bacterial; D009692:Nucleic Acid Heteroduplexes; D012293:Rifampin