02738nas a2200493   4500000000100000008004100001260001300042653003200055653001100087653001100098653001900109653002200128653001300150653001100163653001200174653002500186653001400211653003000225653003600255653002600291100001900317700001300336700001400349700002000363700001500383700001400398700001500412700001600427700001100443700001500454700001700469700001900486700001200505700001200517700001500529700001400544245018300558856004800741300001000789490001600799050001600815520139900831022001402230       2012                            d  c2012 Dec10aBacterial Typing Techniques10aBiopsy10aBrazil10aDNA, Bacterial10aGenetic Variation10agenotype10aHumans10aleprosy10aMycobacterium leprae10aPhylogeny10apolymerase chain reaction10aPolymorphism, Single Nucleotide10aStaining and Labeling1 aBrum Fontes AN1 aGomes HM1 aAraujo MI1 aAlbuquerque ECA1 aBaptista I1 aMoura MMF1 aRezende DS1 aPessolani M1 aLara F1 aPontes MAA1 aGonçalves H1 aLucena-Silva N1 aSarno E1 aVissa V1 aBrennan PJ1 aSuffys PN00aGenotyping of Mycobacterium leprae present on Ziehl-Neelsen-stained microscopic slides and in skin biopsy samples from leprosy patients in different geographic regions of Brazil.  uhttp://www.scielo.br/pdf/mioc/v107s1/21.pdf  a143-90 v107 Suppl 1  aFONTES 20123 a<p>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.</p>  a1678-8060