Article
Genotyping of Mycobacterium leprae present on Ziehl-Neelsen-stained microscopic slides and in skin biopsy samples from leprosy patients in different geographic regions of Brazil
Registro en:
FONTES, Amanda Nogueira Brum; et al. Genotyping of Mycobacterium leprae present on Ziehl-Neelsen-stained microscopic slides and in skin biopsy samples from leprosy patients in different geographic regions of Brazil. Mem Inst Oswaldo Cruz, v.107, Suppl. I, p.143-149, 2012.
0074-0276
1678-8060
Autor
Fontes, Amanda Nogueira Brum
Gomes, Harrison Magdinier
Araujo, Marcelo Ivens de
Albuquerque, Edson Cláudio Araripe de
Baptista, Ida Maria Foschiani Dias
Moura, Maria Manuela da Fonseca
Rezende, Denise Silva
Pessolani, Maria Cristina Vidal
Lara, Flavio Alves
Pontes, Maria Araci de Andrade
Gonçalves, Heitor de Sá
Silva, Norma Lucena
Sarno, Euzenir Nunes
Vissa, Varalakshmi D.
Brennan, Patrick J.
Suffys, Philip Noel
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.