title: Development and evaluation of a line probe assay for rapid identification of pncA mutations in pyrazinamide-resistant mycobacterium tuberculosis strains.
abstract: Resistance of Mycobacterium tuberculosis to pyrazinamide (PZA) derives mainly from mutations in the pncA gene. We developed a reverse hybridization-based line probe assay with oligonucleotide probes designed to detect mutations in pncA. The detection of PZA resistance was evaluated in 258 clinical isolates of M. tuberculosis. The sensitivity and specificity of PZA resistance obtained by this new assay were both 100%, consistent with the results of conventional PZA susceptibility testing. This assay can be used with sputa from tuberculosis patients. It appears to be reliable and widely applicable and, given its simplicity and rapid performance, will be a valuable tool for diagnostic use.
authors.affiliation: Department of Infectious Diseases, Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku, Tokyo 162-8655, Japan.
authors.forename: Jun-Ichiro
authors.initials: J
authors.lastname: Sekiguchi
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Nakamura
authors.forename: Tohru
authors.initials: T
authors.lastname: Miyoshi-Akiyama
authors.forename: Fumiko
authors.initials: F
authors.lastname: Kirikae
authors.forename: Intetsu
authors.initials: I
authors.lastname: Kobayashi
authors.forename: Ewa
authors.initials: E
authors.lastname: Augustynowicz-Kopec
authors.forename: Zofia
authors.initials: Z
authors.lastname: Zwolska
authors.forename: Koji
authors.initials: K
authors.lastname: Morita
authors.forename: Toshinori
authors.initials: T
authors.lastname: Suetake
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yoshida
authors.forename: Seiya
authors.initials: S
authors.lastname: Kato
authors.forename: Toru
authors.initials: T
authors.lastname: Mori
authors.forename: Teruo
authors.initials: T
authors.lastname: Kirikae
mesh_terms: D000581:Amidohydrolases / Q000235:genetics*; D000995:Antitubercular Agents / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D020125:Mutation, Missense; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D011718:Pyrazinamide / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014376:Tuberculosis / Q000382:microbiology
chemical_list: D000995:Antitubercular Agents; D015345:Oligonucleotide Probes; D011718:Pyrazinamide; D000581:Amidohydrolases; C487496:PncA protein, Mycobacterium tuberculosis