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.
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.
Matsuoka
Masanori
M
Leprosy Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Aye
Khin Saw
KS
Department of Medical Research, Yangon, Myanmar.
Kyaw
Kyaw
K
Central Special Skin Clinic, Yangon General Hospital, Yangon, Myanmar.
Tan
Esterlina Virtudes
EV
Leonard Wood Memorial, Cebu, The Philippines.
Balagon
Ma Victoria
MV
Leonard Wood Memorial, Cebu, The Philippines.
Saunderson
Paul
P
Leonard Wood Memorial, Cebu, The Philippines.
Gelber
Robert
R
Leonard Wood Memorial, Cebu, The Philippines.
Makino
Masanao
M
National Leprosy Hospital Okukomyoen, Okayama, Japan.
Nakajima
Chie
C
Department of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
Suzuki
Yasuhiko
Y
Department of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003906:Developing Countries; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D007917:Leprostatic Agents / Q000494:pharmacology*; D007918:Leprosy / Q000188:drug therapy / Q000453:epidemiology / Q000382:microbiology; D009154:Mutation; D002052:Myanmar; D009166:Mycobacterium leprae / Q000187:drug effects* / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010679:Philippines; D015995:Prevalence
D001426:Bacterial Proteins; D007917:Leprostatic Agents