Characterisation of short tandem repeats for genotyping Mycobacterium leprae.
OBJECTIVE
Establish a typing system for Mycobacterium leprae based on polymorphic DNA structures known as short tandem repeats (STR).


METHODS
Assess 16 polymorphic STR for sensitivity, specificity and reproducibility in standard assays using reference strains of M. leprae.


RESULTS
Primers for 16 STR loci were selected based on PCR product size and for their ability to sequence each STR locus from both directions. All primer pairs produced a visible PCR amplicon of appropriate size from PCR reactions containing 10 M. leprae cells. DNA sequences for each STR locus, except (AT) 15, was correctly identified as M. leprae-specific in replicate samples containing 1000 M. leprae using either the forward or reverse PCR primers. Twelve of 13 M. leprae STR loci were stable during passage in heavily infected armadillo tissues over a 5 year and 7 month infection cycle.


CONCLUSIONS
Certain M. leprae STR provide suitable targets for strain typing with the potential for grouping M. leprae with shared genotypes that may prove useful for establishing linkages between leprosy cases within geographical regions.
Gillis
Thomas
T
National Hansen's Disease Programs, Baton Rouge, LA 70803, USA. tgillis@lsu.edu
Vissa
Varalakshmi
V
Matsuoka
Masanori
M
Young
Saroj
S
Richardus
Jan Hendrik
JH
Truman
Richard
R
Hall
Barry
B
Brennan
Patrick
P
D000818:Animals; D001135:Armadillos; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D007918:Leprosy / Q000276:immunology* / Q000382:microbiology; D051379:Mice; D018895:Microsatellite Repeats; D009166:Mycobacterium leprae / Q000235:genetics*; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial