Biomedical Science Letters (대한의생명과학회지)

The Korean Society for Biomedical Laboratory Sciences (대한의생명과학회)
권호 표지

Detection of Mycobacterium leprae by Real-time PCR Targeting Mycobacterium leprae-Specific Repetitive Element Sequence

  • Jin, Hyun-Woo (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Wang, Hye-Young (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Kim, Jong-Pill (Affiliated Hospital, Korean Leprosy Control Association) ;
  • Cho, Sang-Nae (Department of Microbiology, Yonsei University College of Medicine) ;
  • Lee, Hye-Young (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
  • Received : 2010.03.19
  • Accepted : 2010.06.21
  • Published : 2010.06.30
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Abstract

Mycobacterium leprae detection is difficult even with molecular biological techniques due to the low sensitivity of current methodologies. In this report, real-time PCR targeting the M. leprae-specific repetitive element (RLEP) sequence was developed as a new diagnostic tool and evaluated using clinical specimens. For this, M. leprae DNAs were extracted from skin biopsy specimens from 80 patients and analyzed by real-time PCR using TaqMan probe. Then, the detection efficiency of the real-time PCR was compared with that of standard PCR. In brief, the rate of positive detection by the standard PCR and real-time PCR was 32.50% and 66.25%, respectively. The results seemed to clearly show that the TaqMan real-time PCR developed in this study may be a useful tool for sensitive detection of M. leprae from clinical specimens.

Keywords

References

  1. Chae GT, Kim MJ, Kang TJ, Lee SB, Shin HK, Kim JP, Ko YH, Kim SH, Kim NH. DNA-PCR and RT-PCR for the 18-kDa gene of Mycobacterium lepraeto assess the efficacy of multidrug therapy for leprosy. J Med Microbiol. 2002. 51: 417-422. https://doi.org/10.1016/S0167-7012(02)00109-4
  2. De wit MYL, Feber WR, Krieg SR. Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues. J Clin Microbiol. 1991. 29: 906-910.
  3. Donoghue HD, Holton J, Spigelman M. PCR primers that can detect low levels of Mycobacterium leprae DNA. J Med Microbiol. 2001. 50: 177-182. https://doi.org/10.1099/0022-1317-50-2-177
  4. Kampirapap K, Singtham N, Klatser PR, Wiriyawipart S. DNA amplification for detection of leprosy and assessment of efficacy of leprosy chemotherapy. Int J Lepr Other Mycobact Dis. 1998. 66: 16-21.
  5. Kang TJ, Kim SJ, Lee SB, Chae GT, Kim JP. Comparison of Two different PCR amplification products (the 18-kDa protein gene vs. RLEP repetitive sequence) in the diagnosis of Mycobacterium leprae. Clin Exp Dermatol. 2003. 28: 420 -424. https://doi.org/10.1046/j.1365-2230.2003.01300.x
  6. Kesanopoulos K, Tzanakaki G, Levidiotou S, Blackwell C, Kremastinou J. Evaluation of touch-down real-time PCR based on SYBR Green I fluorescent dye for the detection of Neisseria meningitidis in clinical samples. FEMS Immunol Med Microbiol. 2005. 43: 419-424. https://doi.org/10.1016/j.femsim.2004.10.011
  7. Kramme S, Bretzel G, Panning M, Kawuma J, Drosten C. Detection and quantification of Mycobacterium lepraein tissue samples by real-time PCR. Med Microbiol Immunol. 2004. 193: 189 -193. https://doi.org/10.1007/s00430-003-0188-8
  8. Kurabachew M, Wondimu A, Ryon JJ. Reverse transcription-PCR detection of Mycobacterium leprae in clinical specimens. J Clin Microbiol. 1998. 36: 1352-1356.
  9. Martinez AN, Britto CF, Nery JA, Sampaio EP, Jardim MR, Sarno EN, Moraes MO. Evaluation of real-time and conventional PCR targeting complex 85 genes for detection of Mycobacterium leprae DNA in skin biopsy samples from patients diagnosed with leprosy. J Clin Microbiol. 2006. 44: 3154-3159. https://doi.org/10.1128/JCM.02250-05
  10. Patrocinio LG, Goulart IM, Goulart LR, Patrocinio JA, Ferreira FR, Fleury RN. Detection of Mycobacterium leprae in nasal mucosa biopsies by the polymerase chain reaction. FEMS Immunol Med Microbiol. 2005. 44: 311-316. https://doi.org/10.1016/j.femsim.2005.01.002
  11. Phetsuksiri B, Rudeeaneksin J, Supapkul P, Wachapong S, Mahotarn K, Brennan PJ. A simplified reverse transcriptase PCR for rapid detection of Mycobacterium leprae in skin specimens. FEMS Immunol Med Microbiol. 2006. 48: 319 -328. https://doi.org/10.1111/j.1574-695X.2006.00152.x
  12. Rondini S, Mensah-Quainoo E, Troll H, Bodmer T, Pluschke G. Development and application of real-time PCR assay for quantification of Mycobacterium ulcerans DNA. J Clin Microbiol. 2003. 41: 4231-4237. https://doi.org/10.1128/JCM.41.9.4231-4237.2003
  13. Shamsi FA, Chaudhry IA, Moraes MO, Martinez AN, Riley FC. Detection of Mycobacterium lepraein ocular tissues by histopathology and real-time polymerase chain reaction. Ophthalmic Res. 2007. 39: 63-68. https://doi.org/10.1159/000099375
  14. Talhari S. Leprosy diagnosis, classification and prognosis. Int J Lepr Other Mycobact Dis. 1996. 64: S13-S15.
  15. World Health Organization. New case detection trends in leprosy. 2006. Available from:
  16. Yoon KH, Cho SN, Lee MK, Abalos RM, Cellona RV, Fajardo Jr TT, Guido LS, Dela Cruz EC, Walsh GP, Kim JD. Evaluation of polymerase chain reaction amplification of Mycobacterium leprae-specific repetitive sequence in biopsy specimens from leprosy patients. J Clin Microbiol. 1993. 31: 895-899.
  17. Yoon KH, Cho SN, Gerald PW, Lee JB, Kim JD. Detection of Mycobacterium leprae in Skin Biopsy Specimens From Leprosy Patients by Polymerase Chain Reaction. Kor J Dermatol. 1994. 32: 409-415.