title: Detection of Mycobacterium tuberculosis in clinical specimens by polymerase chain reaction and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test.
abstract: The polymerase chain reaction (PCR) using oligonucleotides based on the repetitive sequence (IS986) of Mycobacterium tuberculosis as a primer and the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD), which combines an M. tuberculosis rRNA amplification method with the hybridization protection assay format, were evaluated for detection of M. tuberculosis in clinical samples. The detection limits of these two assay systems based on nucleic acid amplification for cultured M. tuberculosis were less than 10 cells per reaction. A total of 135 sputum specimens were examined by the two assay systems. The PCR and the MTD systems for detection of M. tuberculosis gave overall positivity rates of 84.2% (32 of 38) and 91.9% (34 of 37), respectively, as compared with 71.9% (23 of 32) by smear and 96.9% (31 of 32) by culture in the liquid medium MB-Check. Procedures for sample preparation used in the two methods were different. Although the sensitivities of the PCR and MTD appeared to be similar to that of culture with the MB-Check system, the two methods based on nucleic acid amplification should be very useful for rapid detection of M. tuberculosis infections without the long time required for culture of M. tuberculosis.
authors.affiliation: Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo.
authors.forename: C
authors.initials: C
authors.lastname: Abe
authors.forename: K
authors.initials: K
authors.lastname: Hirano
authors.forename: M
authors.initials: M
authors.lastname: Wada
authors.forename: Y
authors.initials: Y
authors.lastname: Kazumi
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: Y
authors.initials: Y
authors.lastname: Fukasawa
authors.forename: T
authors.initials: T
authors.lastname: Yoshimura
authors.forename: C
authors.initials: C
authors.lastname: Miyagi
authors.forename: S
authors.initials: S
authors.lastname: Goto
mesh_terms: D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D003951:Diagnostic Errors; D005069:Evaluation Studies as Topic; D006801:Humans; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial