原著

生物発光を用いた結核菌の迅速薬剤感受性試験法
(第 IV 報)：改良した ATP 法と参照法との比較

山　崎　利　雄*1　佐　藤　直　樹*2　山　下　研　也*3
岡　沢　　　豊*4　荒　木　　　一*5　三　輪　昭　成*6
　　　　　　　　　　　　　　　　　　　　　　
Reliability of Bioluminescent Assay(ATP Method) for
Testing Antimicrobial Susceptibility of Mycobacterium tuberculosis

Toshio YAMAZAKI*1, Naoki SATO*2, Kenya YAMASHITA*3,
Yutaka OKAZAWA*4, Hajime ARAKI*5 and Akinari MIWA*6

To evaluate the reliability of our previously reported antimicrobial susceptibility test by ATP method, we have compared our ATP method to the reference test methods such as Mycobacteria Growth Indicator Tube(MGIT) method, minimum inhibitory concentration(MIC) method, NCCLS M24-T agar proportion method(M24-T method), and Vite spectrum method. The concentrations of drugs used for the assessment were isoniazid(INH) 0.1mg/ml, rifampicin(RFP) 2.0mg/ml, ethambutol(EB) 2.5mg/ml, streptomycin(SM) 2.0mg/ml, and kanamycin(KM) 5.0mg/ml. When six M. tuberculosis ATCC strains were subjected to 6 independent experiments by using ATP method, highly reproducible results were obtained on the fifth day of the incubation. We examined correlation among ATP method and reference test methods in drug susceptibility testing for 65 clinical isolates of M. tuberculosis. The correlation between ATP method and MGIT-, MIC-, M24-T method were more than 95% for all drugs. When ATP method and Vite spectrum method was compared, the correlation was 87.7% for INH, 98.5% for RFP, 90.8% for EB, 92.3% for SM, 96.9% for KM. The culture period for determining susceptibility between ATP method and MGIT method was compared by using ATCC reference strains and clinical isolates. Six M. tuberculosis ATCC strains were subjected to 6 independent experiments. By the MGIT method, 8 days were required to obtain the results, whereas 3 days were enough by the ATP method. For 65 clinical isolates, the MGIT method required 9 days for determining susceptibility of all isolates. The ATP method required only 5 days for the same strains. These data demonstrate that the improved ATP method that we reported, is simple, rapid, highly reproducible and nonradiometric, and could be used for the assessment of drug susceptibility for M. tuberculosis with high reliability.
[Rinsho Byori 51 : 194〜200, 2003]

*1Department of Bacteriology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640

【Key Words】Mycobacterium tuberculosis(結核菌)，antimicrobial susceptibility test(薬剤感受性試験)，adeno-sine triphosphate(アデノシン三リン酸)，bioluminescent assay(生物発光法)，filamentous cell treatment：FCT
　　　　　　　　　　　　　　　　　　　　　
受付2002年10月4日・受理2003年2月13日
*1国立感染症研究所細菌第一部(〒162-8640 東京都新宿区戸山1-23-1)
*2〜6極東製薬工業�轄ｔ牛H場研究開発部(〒318-0004 高萩市大字上手綱字朝山3333-26)

E-mail :toshiyam@nih.go.jp