This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by de Wit, M Y Articles by Hartskeerl, R A Search for Related Content PubMed PubMed Citation Articles by de Wit, M Y Articles by Hartskeerl, R A

Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues.

N. H. Swellengrebel Laboratory of Tropical Hygiene, Royal Tropical Institute, Amsterdam, The Netherlands.

ABSTRACT

The polymerase chain reaction (PCR) based on the selective amplification of a 530-bp fragment of the gene encoding the proline-rich antigen of Mycobacterium leprae was applied on sections of fixed or frozen biopsy samples from leprosy patients. A simple procedure for the extraction of DNA from M. leprae in clinical specimens that provided suitable template DNA for amplification was developed. When PCR was applied on frozen sections, positive amplification in samples from all untreated acid-fast bacillus (AFB)-positive patients and in samples from 56% of the untreated AFB-negative patients could be detected, while biopsy samples from patients with skin diseases other than leprosy were all PCR negative. With neutral Formalin-fixed biopsy samples, positive amplification in 92% of the samples from untreated AFB-positive patients and in 61% of the samples from untreated AFB-negative patients could be detected by PCR. Biopsy samples exposed to mercuric chloride or nonbuffered formaldehyde containing fixatives were not suitable for application of PCR. This PCR holds promise as a tool for studies on M. leprae infection.

This article has been cited by other articles:

• Scollard, D. M., Adams, L. B., Gillis, T. P., Krahenbuhl, J. L., Truman, R. W., Williams, D. L. (2006). The Continuing Challenges of Leprosy. Clin. Microbiol. Rev. 19: 338-381 [Abstract] [Full Text]  
• Matsuoka, M., Zhang, L., Budiawan, T., Saeki, K., Izumi, S. (2004). Genotyping of Mycobacterium leprae on the Basis of the Polymorphism of TTC Repeats for Analysis of Leprosy Transmission. J. Clin. Microbiol. 42: 741-745 [Abstract] [Full Text]  
• Stienstra, Y., van der Werf, T. S., Guarner, J., Raghunathan, P. L., Spotts Whitney, E. A., van der Graaf, W. T. A., Asamoa, K., Tappero, J. W., Ashford, D. A., King, C. H. (2003). Analysis of an IS2404-Based Nested PCR for Diagnosis of Buruli Ulcer Disease in Regions of Ghana Where the Disease Is Endemic. J. Clin. Microbiol. 41: 794-797 [Abstract] [Full Text]  
• CHAE, G.-T., KIM, M.-J., KANG, T.-J., LEE, S.-B., SHIN, H.-K., KIM, J.-P., KO, Y.-H., KIM, S.-H., KIM, N.-H. (2002). DNA-PCR and RT-PCR for the 18-kDa gene of Mycobacterium leprae to assess the efficacy of multi-drug therapy for leprosy. J Med Microbiol 51: 417-422 [Abstract] [Full Text]  
• Maeda, S., Matsuoka, M., Nakata, N., Kai, M., Maeda, Y., Hashimoto, K., Kimura, H., Kobayashi, K., Kashiwabara, Y. (2001). Multidrug Resistant Mycobacterium leprae from Patients with Leprosy. Antimicrob. Agents Chemother. 45: 3635-3639 [Abstract] [Full Text]  
• Kurabachew, M., Wondimu, A., Ryon, J. J. (1998). Reverse Transcription-PCR Detection of Mycobacterium leprae in Clinical Specimens. J. Clin. Microbiol. 36: 1352-1356 [Abstract] [Full Text]