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J Med Microbiol 57 (2008), 1213-1219; DOI: 10.1099/jmm.0.2008/002600-0
© 2008 Society for General Microbiology
ISSN 1473-5644

A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries

Masanori Matsuoka1,{dagger}, Khin Saw Aye2, Kyaw Kyaw3, Esterlina Virtudes Tan4, Ma Victoria Balagon4, Paul Saunderson4, Robert Gelber4, Masanao Makino5, Chie Nakajima6 and Yasuhiko Suzuki6,{dagger}

1 Leprosy Research Center, National Institute of Infectious Diseases, Tokyo, Japan

2 Department of Medical Research, Yangon, Myanmar

3 Central Special Skin Clinic, Yangon General Hospital, Yangon, Myanmar

4 Leonard Wood Memorial, Cebu, The Philippines

5 National Leprosy Hospital Okukomyoen, Okayama, Japan

6 Department of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan

Correspondence
Masanori Matsuoka
matsuoka{at}nih.go.jp

Received 14 April 2008
Accepted 18 June 2008

A simple method to detect mutations in the genome of Mycobacterium leprae that confer resistance to key drugs for leprosy was exploited on the basis of a reverse hybridization system. A series of oligonucleotide probes corresponding to each mutation in the folP1, rpoB and gyrA genes for dapsone, rifampicin and ofloxacin resistance, respectively, were selected and fixed on a glass slide as capture probes, to develop a DNA microarray termed the leprosy drug susceptibility-DNA microarray (LDS-DA). Mutations in clinical isolates of M. leprae were successfully identified by the LDS-DA. Feasibility studies were conducted to evaluate the performance of the LDS-DA in two developing countries, Myanmar and the Philippines. The high concordance of the results obtained by this method with the results of nucleotide sequencing strongly supports the applicability of the LDS-DA as a drug susceptibility test in place of sequencing, a time-consuming and costly procedure. This is a rapid and simple method for the simultaneous susceptibility testing of three front-line drugs for leprosy, and solves the problems of previously reported methods.

Abbreviations: BI, bacterial index; DRDR, drug-resistance-determining region; LDS-DA, leprosy drug susceptibility-DNA microarray; MDT, multidrug therapy.

{dagger}These authors contributed equally to this work.






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