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Journal of Clinical Microbiology, September 2007, p. 2802-2807, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00352-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development and Evaluation of a Line Probe Assay for Rapid Identification of pncA Mutations in Pyrazinamide-Resistant Mycobacterium tuberculosis Strains{triangledown}

Jun-Ichiro Sekiguchi,1 Tomohiko Nakamura,2 Tohru Miyoshi-Akiyama,1 Fumiko Kirikae,1 Intetsu Kobayashi,3 Ewa Augustynowicz-Kopec,4 Zofia Zwolska,4 Koji Morita,5 Toshinori Suetake,2 Hiroshi Yoshida,2 Seiya Kato,6 Toru Mori,7 and Teruo Kirikae1*

Department of Infectious Diseases, Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku, Tokyo 162-8655, Japan,1 Third Department, Research and Development Laboratory, Nipro Corporation, 3023 Noji, Kusatsu, Shiga 525-0055, Japan,2 Mitsubishi Chemical Medience Corporation, 3-30-1 Shimura, Itabashi, Tokyo 174-8555, Japan,3 National Research Institute of Tuberculosis and Lung Diseases, Plocka St. 26, Warsaw 01-138, Poland,4 Department of Microbiology, Kyorin University School of Health Sciences, 476 Miyashita, Hachioji, Tokyo 192-8508, Japan,5 Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Matsuyama 3-1-24, Kiyose, Tokyo 204-8533, Japan,6 Leprosy Research Center, National Institute of Infectious Diseases, Aoba 4-2-1, Higashimurayama, Tokyo 189-0002, Japan7

Received 14 February 2007/ Returned for modification 28 March 2007/ Accepted 19 June 2007

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.

* Corresponding author. Mailing address: Department of Infectious Diseases, Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku, Tokyo 162-8655, Japan. Phone: (81) 3 3202 7181, ext. 2838. Fax: (81) 3 3202 7364. E-mail: tkirikae{at}ri.imcj.go.jp

{triangledown} Published ahead of print on 27 June 2007.

Journal of Clinical Microbiology, September 2007, p. 2802-2807, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00352-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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