RESUMEN
We analysed 16 variable number tandem repeats (VNTR) and three single-nucleotide polymorphisms (SNP) in Mycobacterium leprae present on 115 Ziehl-Neelsen (Z-N)-stained slides and in 51 skin biopsy samples derived from leprosy patients from Ceará (n = 23), Pernambuco (n = 41), Rio de Janeiro (n = 22) and Rondônia (RO) (n = 78). All skin biopsies yielded SNP-based genotypes, while 48 of the samples (94.1%) yielded complete VNTR genotypes. We evaluated two procedures for extracting M. leprae DNA from Z-N-stained slides: the first including Chelex and the other combining proteinase and sodium dodecyl sulfate. Of the 76 samples processed using the first procedure, 30.2% were positive for 16 or 15 VNTRs, whereas of the 39 samples processed using the second procedure, 28.2% yielded genotypes defined by at least 10 VNTRs. Combined VNTR and SNP analysis revealed large variability in genotypes, but a high prevalence of SNP genotype 4 in the Northeast Region of Brazil. Our observation of two samples from RO with an identical genotype and seven groups with similar genotypes, including four derived from residents of the same state or region, suggest a tendency to form groups according to the origin of the isolates. This study demonstrates the existence of geographically related M. leprae genotypes and that Z-N-stained slides are an alternative source for M. leprae genotyping.
Asunto(s)
ADN Bacteriano/análisis , Variación Genética , Lepra/microbiología , Mycobacterium leprae/genética , Técnicas de Tipificación Bacteriana , Biopsia , Brasil , Genotipo , Humanos , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido Simple , Coloración y EtiquetadoRESUMEN
We analysed 16 variable number tandem repeats (VNTR) and three single-nucleotide polymorphisms (SNP) in Mycobacterium leprae present on 115 Ziehl-Neelsen (Z-N)-stained slides and in 51 skin biopsy samples derived from leprosy patients from Ceará (n = 23), Pernambuco (n = 41), Rio de Janeiro (n = 22) and Rondônia (RO) (n = 78). All skin biopsies yielded SNP-based genotypes, while 48 of the samples (94.1%) yielded complete VNTR genotypes. We evaluated two procedures for extracting M. leprae DNA from Z-N-stained slides: the first including Chelex and the other combining proteinase and sodium dodecyl sulfate. Of the 76 samples processed using the first procedure, 30.2% were positive for 16 or 15 VNTRs, whereas of the 39 samples processed using the second procedure, 28.2% yielded genotypes defined by at least 10 VNTRs. Combined VNTR and SNP analysis revealed large variability in genotypes, but a high prevalence of SNP genotype 4 in the Northeast Region of Brazil. Our observation of two samples from RO with an identical genotype and seven groups with similar genotypes, including four derived from residents of the same state or region, suggest a tendency to form groups according to the origin of the isolates. This study demonstrates the existence of geographically related M. leprae genotypes and that Z-N-stained slides are an alternative source for M. leprae genotyping.
Asunto(s)
Humanos , ADN Bacteriano/análisis , Variación Genética , Lepra/microbiología , Mycobacterium leprae/genética , Técnicas de Tipificación Bacteriana , Biopsia , Brasil , Genotipo , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido Simple , Coloración y EtiquetadoRESUMEN
Fundamentos: A integração das artes de controle da hanseníase, as estratégias dos Programas de Saúde da Família e Agentes Comunitários de Saúde, na conformidade do processo de reorganização da atenção básica, fundamentou a atualização das normas da legislação sobre o controle da doença no Brasil. A classificação operacional adotada, para aloação do paciente na polioquimioterapia, foi adaptada da sugerida pela Organização Mundial da Saúde, essencialmente clínica, baseada no número de lesões cutaneas