title: Routine use of a simple low-cost genotypic assay for the identification of mycobacteria in a high throughput laboratory.
abstract: A novel polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene was used for the routine identification of mycobacteria in a high throughput clinical laboratory. A total of 2036 clinical isolates were tested by PRA in conjunction with other methods. The PRA identification of M. tuberculosis complex was 100% sensitive and specific, and 74.5% of nontuberculous mycobacteria (NTM) were correctly identified. It gave highly consistent results for Mycobacterium avium complex (MAC) species and for most isolates of M. fortuitum, M. chelonae, and M. kansasii. It had proven to be highly robust and stable despite usage on such a large-scale and is thus particularly suitable for use in high throughput laboratories in areas with a high incidence of tuberculosis.
authors.affiliation: TB Reference Laboratory, Pathology Service, Department of Health, Hong Kong, China.
authors.forename: Derek A
authors.initials: DA
authors.lastname: Wong
authors.forename: Peter C W
authors.initials: PC
authors.lastname: Yip
authors.forename: Denise L W
authors.initials: DL
authors.lastname: Tse
authors.forename: Viola W N
authors.initials: VW
authors.lastname: Tung
authors.forename: Danny T L
authors.initials: DT
authors.lastname: Cheung
authors.forename: Kai Man
authors.initials: KM
authors.lastname: Kam
mesh_terms: D000465:Algorithms; D001426:Bacterial Proteins; D015373:Bacterial Typing Techniques; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D005838:Genotype; D006801:Humans; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011786:Quality Control; D015203:Reproducibility of Results
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; C074119:heat-shock protein 65, Mycobacterium; D018833:Chaperonins