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Journal of Clinical Microbiology, April 2004, p. 1666-1672, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1666-1672.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Multiple Polymorphic Loci for Molecular Typing of Strains of Mycobacterium leprae

Nathan A. Groathouse,1 Becky Rivoire,1 Hansuk Kim,2 Hyeyoung Lee,2 Sang-Nae Cho,3 Patrick J. Brennan,1 and Varalakshmi D. Vissa1*

Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523-1682,1 Department of Biomedical Laboratory Science, College of Health Science,2 Department of Microbiology, College of Medicine, Yonsei University, Seoul, Republic of Korea3

Received 14 September 2003/ Returned for modification 25 November 2003/ Accepted 15 December 2003

The need for molecular tools for the differentiation of isolates of Mycobacterium leprae, the organism that causes leprosy, is urgent in view of the continuing high levels of new case detection, despite years of aggressive chemotherapy and the consequent reduction in the prevalence of leprosy. The slow onset of leprosy and the reliance on physical examination for detection of disease have restricted the epidemiological tracking necessary to understand and control transmission. Two genetic loci in several isolates of M. leprae have previously been demonstrated to contain variable-number tandem repeats (VNTRs). On the basis of these reports and the availability of the full genome sequence, multiple-locus VNTR analysis for strain typing has been undertaken. A panel of 11 short tandem repeat (STR) loci with repeat units of 1, 2, 3, 6, 12, 18, 21, and 27 bp from four clinical isolates of M. leprae propagated in armadillo hosts were screened by PCR. Fragment length polymorphisms were detected at 9 of the 11 loci by agarose gel electrophoresis. Sequencing of representative DNA products confirmed the presence of VNTRs between isolates. The application of nine new polymorphic STRs in conjunction with automated methods for electrophoresis and size determination allows greater discrimination between isolates of M. leprae and enhances the potential of this technique to track the transmission of leprosy.

* Corresponding author. Mailing address: Department of Microbiology, Immunology and Pathology, Campus Delivery 1682, Colorado State University, Fort Collins, CO 80523-1682. Phone: (970) 491-3525. Fax: (970) 491-1815. E-mail: Varalakshmi.Vissa{at}ColoState.edu.

Journal of Clinical Microbiology, April 2004, p. 1666-1672, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1666-1672.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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