Skip to content
1887

Abstract

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 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.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47499-0
2008-04-01
2025-12-06

Metrics

Loading full text...

Full text loading...

References

  1. Boehme C. C., Nabeta P., Henostroza G., Raqib R., Rahim Z., Gerhardt M., Sanga E., Hoelscher M., Notomi T. other authors 2007; Operational feasibility of using loop-mediated isothermal amplification for diagnosis of pulmonary tuberculosis in microscopy centers of developing countries. J Clin Microbiol 45:1936–1940 [CrossRef]
    [Google Scholar]
  2. Centers for Disease Control and Prevention; 1992; National action plan to combat multidrug-resistant tuberculosis: summary of a conference; management of persons exposed to multidrug-resistant tuberculosis. Morbid Mortal Weekly Rep 41:5–48
    [Google Scholar]
  3. Iwamoto T., Sonobe T., Hayashi K. 2003; Loop-mediated isothermal amplification for direct detection of Mycobacterium tuberculosis complex, M. avium , and M. intracellulare . J Clin Microbiol 41:2616–2622 [CrossRef]
    [Google Scholar]
  4. Jonas V., Alden M. J., Curry J. I., Kamisango K., Knott C. A., Lankford R., Wolfe J. M., Moore D. F. 1993; Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by amplification of rRNA. J Clin Microbiol 31:2410–2416
    [Google Scholar]
  5. Kent B. D., Kubica G. P. 1985 Public Health Mycobacteriology: a Guide for the Level III Laboratory Atlanta: US Department of Health and Human Services, Centers for Disease Control;
    [Google Scholar]
  6. Kimura H., Ihira M., Enomoto Y., Kawada J., Ito J., Morishima T., Yoshikawa T., Asano Y. 2005; Rapid detection of herpes simplex virus DNA in cerebrospinal fluid: comparison between loop-mediated isothermal amplification and real-time PCR. Med Microbiol Immunol 194:181–185 [CrossRef]
    [Google Scholar]
  7. Kuboki N., Inoue N., Sakurai T., Cello F. D., Grab D. J., Suzuki H., Sugimoto C., Igarashi I. 2003; Loop-mediated isothermal amplification for detection of African trypanosomes. J Clin Microbiol 41:5517–5524 [CrossRef]
    [Google Scholar]
  8. Notomi T., Okayama H., Masubuchi H., Yonekawa T., Watanabe K., Amino N. 2000; Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63 [CrossRef]
    [Google Scholar]
  9. NTP 2004 Annual report of National Tuberculosis Control Programme Nepal His Majesty's Government of Nepal, Ministry of Health, Department of Health Services;
    [Google Scholar]
  10. Parida M., Inoue G. P. S., Hasebe F., Morita K. 2004; Real-time reverse transcription loop-mediated isothermal amplification for rapid detection of West Nile virus. J Clin Microbiol 42:257–263 [CrossRef]
    [Google Scholar]
  11. Parida M., Horioke K., Ishida H., Dash P. K., Saxena P., Jana A. M., Islam M. A., Inoue S., Hosaka N., Morita K. 2005; Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loop-mediated isothermal amplification assay. J Clin Microbiol 43:2895–2903 [CrossRef]
    [Google Scholar]
  12. Ruiz-Serrano M. J., Albadalejo J., Martinez-Sanchez L., Bouza E. 1998; LCx: a diagnostic alternative for the early detection of Mycobacterium tuberculosis complex. Diagn Microbiol Infect Dis 32:259–264 [CrossRef]
    [Google Scholar]
  13. Suzuki Y., Katsukawa C., Inoue K., Yin Y. P., Tasaka H., Ueba N., Makino M. 1995; Mutations in rpoB gene of rifampicin resistant clinical isolates of Mycobacterium tuberculosis in Japan. Kansenshogaku Zasshi 69:413–419 [CrossRef]
    [Google Scholar]
  14. Takakura S., Tsuchiya S., Isawa Y., Yasukawa K., Hayashi T., Tomita M., Suzuki K., Hasegawa T., Tagami T. other authors 2005; Rapid detection of Mycobacterium tuberculosis in respiratory samples by transcription-reverse transcription concerted reaction with an automated system. J Clin Microbiol 43:5435–5439 [CrossRef]
    [Google Scholar]
  15. Truant J. P., Brett W. A., Thomas W. Jr 1962; Fluorescence microscopy of tubercle bacilli stained with auramine and rhodamine. Henry Ford Hosp Med Bull 10:287–296
    [Google Scholar]
  16. WHO 2007 Global Tuberculosis Control: Surveillance, Planning, Financing . WHO report (document WHO/HTM/TB/2005, 349) Geneva: World Health Organization;
    [Google Scholar]
  17. Woods S. A., Cole S. T. 1989; A rapid method for the detection of potentially viable Mycobacterium leprae in human biopsies: a novel application of PCR. FEMS Microbiol Lett 53:305–309
    [Google Scholar]
  18. Yoda T., Suzuki Y., Yamazaki K., Sakon N., Kanki M., Aoyama I., Tsukamoto T. 2007; Evaluation and application of reverse transcription loop-mediated isothermal amplification for detection of noroviruses. J Med Virol 79:326–334 [CrossRef]
    [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.47499-0
Loading
/content/journal/jmm/10.1099/jmm.0.47499-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An error occurred
Approval was partially successful, following selected items could not be processed due to error