Título:	Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil.
Autoria:	SILVA, M. R. GUIMARÃES, M. D. C. VEIGA, V. M. de O. MOREIRA, A. dos S. COSTA, R. R. da ABI-ZAID, K. C. F. ROCHA, A. da S. SUFFYS, P. N.
Afiliação:	MARCIO ROBERTO SILVA, CNPGL; MARK DREW CROSLAND GUIMARÃES, UFMG; VANIA MARIA DE OLIVEIRA VEIGA, CNPGL; ALINE DOS SANTOS MOREIRA, Laboratório de Genômica Funcional e Bioinformática; RONALDO RODRIGUES DA COSTA, UFJF; FHEMIG; KELLY CRISTINA FERREIRA ABI-ZAID, FHEMIG; ADALGIZA DA SILVA ROCHA, FIOCRUZ; PHILIP NOEL SUFFYS, FIOCRUZ.
Ano de publicação:	2011
Referência:	Memorias do Instituto Oswaldo Cruz, v. 106, n. 1, p. 9-15, 2011.
Conteúdo:	A cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC.
Thesagro:	Mycobacterium Bovis
NAL Thesaurus:	Mycobacterium tuberculosis polymerase chain reaction
Palavras-chave:	OxyR pseudogene Sputum smear Molecular sequence data
Digital Object Identifier:	https://doi.org/10.1590/S0074-02762011000100002
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CNPGL)