PCR-restriction fragment length polymorphism analysis as a tool for Mycobacterium species identification in lepromas for lepromin production.
OBJECTIVE
The aim of the present work was to standardise a PCR-Restriction Fragment Length Polymorphism analysis (PRA) as a tool to detect the mycobacteriologic composition of lepromas from leprosy patients used in the production of lepromin to improve the quality of the Mitsuda test.


METHODS
PCR-Restriction Fragment Length Polymorphism analysis using hsp65 and rpoB genes were applied to 11 reference strains of mycobacteria, including M. leprae, and the obtained PRA profiles were compared to mycobacteria in clinical specimens.


RESULTS
Out of the biopsies studied, 522% had DNA fragment amplified for both genes (hsp65 and rpoB) for M. leprae. However, other Mycobacterium species were observed in samples of lepromatous leprosy patients. Here we discussed the importance of mycobacteria identification in the antigen of Mitsuda production to be used in the evaluation of leprosy.


CONCLUSIONS
Our results suggest that the use of the molecular approach for sample selection can contribute to an improvement in the quality of produced lepromin.
Alban
Silvana M
SM
Federal University of Paraná, 81531-990 Curitiba, PR, Brazil.
Sella
Sandra R B R
SR
Miranda
Ruy N
RN
Mira
Marcelo Távora
MT
Soccol
Vanete Thomaz
VT
D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods / Q000592:standards; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods* / Q000592:standards; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005838:Genotype; D006801:Humans; D007916:Lepromin / Q000302:isolation & purification*; D007918:Leprosy / Q000382:microbiology* / Q000473:pathology; D009166:Mycobacterium leprae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012150:Polymorphism, Restriction Fragment Length
D001426:Bacterial Proteins; D018834:Chaperonin 60; C074119:heat-shock protein 65, Mycobacterium; D007916:Lepromin; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D018833:Chaperonins