期刊论文详细信息
Memórias do Instituto Oswaldo Cruz
Plasmid-based controls to detect rpoB mutations in Mycobacterium tuberculosis by quantitative polymerase chain reaction-high-resolution melting
Joas Lucas Da Silva1  Gabriela Guimaraes Sousa Leite1  Gisele Medeiros Bastos1  Beatriz Cacciacarro Lucas1  Daniel Keniti Shinohara1  Joice Sayuri Takinami1  Marcelo Miyata1  Cristina Moreno Fajardo1  André Ducati Luchessi1  Clarice Queico Fujimura Leite2  Rosilene Fressatti Cardoso2  Rosario Dominguez Crespo Hirata1  Mario Hiroyuki Hirata1 
[1] ,Universidade de São Paulo Faculdade de Ciências Farmacêuticas São Paulo SP ,Brasil
关键词: drug resistance;    rifampicin;    Mycobacterium tuberculosis;   
DOI  :  10.1590/S0074-02762013000100017
来源: SciELO
PDF
【 摘 要 】

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.

【 授权许可】

CC BY   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License

【 预 览 】
附件列表
Files Size Format View
RO202005130048346ZK.pdf 579KB PDF download
  文献评价指标  
  下载次数:9次 浏览次数:8次