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Journal of Clinical Microbiology, November 2005, p. 5435-5439, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5435-5439.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Mycobacterium tuberculosis in Respiratory Samples by Transcription-Reverse Transcription Concerted Reaction with an Automated System

Shunji Takakura,1* Shigeo Tsuchiya,2 Yuichi Isawa,2 Kiyoshi Yasukawa,2,{dagger} Toshinori Hayashi,2 Motohisa Tomita,3 Katsuhiro Suzuki,3 Tatsuro Hasegawa,4 Takanori Tagami,4 Atsuyuki Kurashima,4 and Satoshi Ichiyama1

Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, 54 Shogoin-Kawaharacho, Sakyo-ku, Kyoto 6068507, Japan,1 Scientific Instruments Division, Tosoh Corporation, 2743-1 Hayakawa, Ayase, Kanagawa 2521123, Japan,2 Clinical Research Center, Kinki-chuo Chest Medical Center, 1180 Nagasone-cho, Sakai, Osaka 5918555, Japan,3 National Tokyo Hospital, 3-1-1 Takeoka, Kiyoshe-shi, Tokyo 2048585, Japan4

Received 6 June 2005/ Returned for modification 28 June 2005/ Accepted 10 August 2005

The aim of this study was to evaluate the performance of the transcription-reverse transcription concerted (TRC) method for the detection of Mycobacterium tuberculosis complex (MTC) 16S rRNA in clinical respiratory samples for the diagnosis of pulmonary tuberculosis. TRC is a novel method that enables the rapid and the completely homogeneous real-time monitoring of isothermal sequence RNA amplification without any postamplification procedure. The detection limit of the TRC method for MTC was one organism per 100 µl of sputum. The specificity of the method was confirmed by the absence of positive signals for sputum containing 106 M. avium or M. kansasii organisms per 100 µl. A total of 201 respiratory samples from patients diagnosed with or suspected of having tuberculosis were tested. Of the 72 MTC culture-positive samples, the TRC method was positive for 52 (sensitivity, 72.2%), whereas the Roche COBAS AMPLICOR PCR was positive for 58 (sensitivity, 80.6%). Both the TRC method and the COBAS AMPLICOR PCR showed no positive identification for any of the 129 culture-negative samples. The percent agreement between the two methods was 95% (191 of 201 samples). The high sensitivity and specificity together with shorter detection time (within 1 h) of the TRC method allow it to be proposed as a useful method for the rapid detection of MTC in respiratory samples.

* Corresponding author. Mailing address: Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, 54 Shogoin-Kawaharacho, Sakyo-ku, Kyoto 6068507, Japan. Phone: 81-75-751-3503. Fax: 81-75-751-3233. E-mail: stakakr{at}kuhp.kyoto-u.ac.jp.

{dagger} Present address: Department of Food Biology, Kyoto University Graduate School of Agricultural Science, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto 6068502, Japan.

Journal of Clinical Microbiology, November 2005, p. 5435-5439, Vol. 43, No. 11
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.11.5435-5439.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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