title: Evaluation of polymerase chain reaction and restriction enzyme analysis for routine identification of mycobacteria: accuracy, rapidity, and cost analysis.
abstract: Polymerase chain reaction and restriction enzyme analysis (PCR-REA) of the hsp65 gene was evaluated for use as a routine identification method for identifying mycobacteria. The accuracy, rapidity, and cost were assessed compared with the conventional biochemical method. Five hundred and forty-one mycobacterial clinical isolates obtained from the Department of Microbiology, Faculty of Medicine at Siriraj Hospital, Mahidol University, were submitted for PCR-REA and biochemical identification. PCR-REA showed high concordant result with 100, 96.2, and 94.1% for identification of Mycobacterium tuberculosis, rapid- and slow-growing mycobacteria, respectively. Discordant results were obtained from 24 (4.4%) out of 541 isolates, consisting of 9 rapid growers (6 M. chelonae, 2 M. abscessus, and 1 M. fortuitum) and 15 slow growers (9 M. scrofulaceum, 2 M. gordonae, 1 M. avium, 1 M. kansasii, 1 M. malmoense, and 1 M. terrae complex). PCR-REA demonstrated not only accurate results but was also less expensive (2.1 US dollars/sample). This method was rapid with a turn-around time of 30 hours compared with 2-4 weeks for the conventional method.
authors.affiliation: Department of Microbiology, Faculty of Medicine at Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Therdsak
authors.initials: T
authors.lastname: Prammananan
authors.forename: Wattana
authors.initials: W
authors.lastname: Cheunoy
authors.forename: Preeyawit
authors.initials: P
authors.lastname: Na-Ubol
authors.forename: Nipa
authors.initials: N
authors.lastname: Tingtoy
authors.forename: Somboon
authors.initials: S
authors.lastname: Srimuang
authors.forename: Angkana
authors.initials: A
authors.lastname: Chaiprasert
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification; D001483:Base Sequence; D018834:Chaperonin 60; D018833:Chaperonins / Q000302:isolation & purification; D003365:Costs and Cost Analysis; D017931:DNA Primers; D004269:DNA, Bacterial; D006801:Humans; D009161:Mycobacterium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D000091402:Prohibitins; D015203:Reproducibility of Results; D015183:Restriction Mapping; D013785:Thailand
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D017931:DNA Primers; D004269:DNA, Bacterial; C000717331:PHB2 protein, human; D000091402:Prohibitins; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins