Please use this identifier to cite or link to this item: http://www.repositorio.ufc.br/handle/riufc/6938
Title in Portuguese: Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting
Author: Silva, Joas Lucas da
Leite, Gabriela Guimaraes Sousa
Bastos, Gisele Medeiros
Lucas, Beatriz Cacciacarro
Shinohara, Daniel Keniti
Takinami, Joice Sayuri
Miyata, Marcelo
Fajardo, Cristina Moreno
Luchessi, André Ducati
Leite, Clarice Queico Fujimura
Cardoso, Rosilene Fressatti
Hirata, Rosario Dominguez Crespo
Hirata, Mario Hiroyuki
Keywords: Rifampina
Mycobacterium tuberculosis
Issue Date: Feb-2013
Publisher: Memórias do Instituto Oswaldo Cruz
Citation: SILVA, J. L. et al. Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting. Mem. Inst. Oswaldo Cruz, Rio de Janeiro, RJ, v. 108, n. 1, p. 106-109, fev. 2013.
Abstract: Quantitative polymerase chain reaction-high-resolution melting (qPCR-HRM) analysis was used to screen for mutations related to drug resistance in Mycobacterium tuberculosis. We detected the C526T and C531T mutations in the rifampicin resistance-determining region (RRDR) of the rpoB gene with qPCR-HRM using plasmid-based controls. A segment of the RRDR region from M. tuberculosis H37Rv and from strains carrying C531T or C526T mutations in the rpoB were cloned into pGEM-T vector and these vectors were used as controls in the qPCR-HRM analysis of 54 M. tuberculosis strains. The results were confirmed by DNA sequencing and showed that recombinant plasmids can replace genomic DNA as controls in the qPCR-HRM assay. Plasmids can be handled outside of biosafety level 3 facilities, reducing the risk of contamination and the cost of the assay. Plasmids have a high stability, are normally maintained in Escherichia coli and can be extracted in large amounts.
URI: http://www.repositorio.ufc.br/handle/riufc/6938
ISSN: 0074-0276
Appears in Collections:DFAR - Artigos publicados em revistas científicas

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