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J Med Microbiol 57 (2008), 439-443; DOI: 10.1099/jmm.0.47499-0
© 2008 Society for General Microbiology
ISSN 1473-5644

Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients

Basu Dev Pandey1,2, Ajay Poudel3, Tomoko Yoda4, Aki Tamaru4, Naozumi Oda5, Yukari Fukushima6, Binod Lekhak3, Basista Risal3, Bishnu Acharya2, Bishwa Sapkota7, Chie Nakajima6, Tooru Taniguchi8, Benjawan Phetsuksiri9 and Yasuhiko Suzuki6

1 Sukraraj Tropical and Infectious Disease Hospital, Kathmandu, Nepal

2 Everest International Clinic and Research Center, Kathmandu, Nepal

3 Department of Microbiology, Tribhuban University, Kathmandu, Nepal

4 Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan

5 Asahi Kasei Co. Ltd, Shizuoka, Japan

6 Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan

7 Department of Research and Biotechnology, Anandaban Hospital, Kathmandu, Nepal

8 Section of Bacterial Infections, Research Collaboration Center on Emerging and Re-emerging Infections, Osaka University, Nonthaburi, Thailand

9 National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand

Correspondence
Yasuhiko Suzuki
suzuki{at}czc.hokudai.ac.jp

Received 8 July 2007
Accepted 9 December 2007

A number of nucleic acid amplification assays (NAAs) have been employed to detect tubercle bacilli in clinical specimens for tuberculosis (TB) diagnosis. Among these, loop-mediated isothermal amplification (LAMP) is an NAA possessing superior isothermal reaction characteristics. In the present study, a set of six specific primers targeting the Mycobacterium tuberculosis 16S rRNA gene with high sensitivity was selected and a LAMP system (MTB-LAMP) was developed. Using this system, a total of 200 sputum samples from Nepalese patients were investigated. The sensitivity of MTB-LAMP in culture-positive samples was 100 % (96/96), and the specificity in culture-negative samples was 94.2 % (98/104, 95 % confidence interval 90.5–97.9 %). The positive and negative predictive values of MTB-LAMP were 94.1 and 100 %, respectively. These results indicate that this MTB-LAMP method may prove to be a powerful tool for the early diagnosis of TB.

Abbreviations: AFB, acid-fast bacilli; LAMP, loop-mediated isothermal amplification; NAA, nucleic acid amplification assay; TB, tuberculosis.






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