Evaluation of six different DNA extraction methods for detection of Mycobacterium tuberculosis by means of PCR-IS6110: preliminary study

BMC Research Notes

Silvana Spindola de Miranda³  Elis Regina Dalla Costa¹  Maria Lúcia Rossetti¹  Wânia da Silva Carvalho²  Isabela Neves de Almeida² 

[1] State Foundation for Production and Research in Health (FEPPS), Porto Alegre, Brazil;Federal University of Minas Gerais, Belo Horizonte, Brazil;Research Group Coordinator FM/UFMG_REDE-TB, Belo Horizonte, Minas Gerais, Brazil

关键词: PCR;    DNA extraction;    IS6110;    M. tuberculosis;   

Others  :  1135078
DOI  :  10.1186/1756-0500-6-561

 received in 2013-05-30, accepted in 2013-12-16,  发布年份 2013

【 摘 要 】

Background

Developments in molecular detection and strain differentiation of members of Mycobacterium tuberculosis complex have proved to be useful. The DNA extraction method influences the amplification efficiency, causing interference on the sensitivity and respective inhibitors. The aim of this study was to standardize a simple and fast DNA extraction method, providing DNA amplification by IS6110-PCR effectively free from undue interferences.

Findings

The efficiency of the six different protocols tested in M. tuberculosis cultures has varied from 75% to 92.5%. This preliminary study evaluating the IS6110 PCR sensitivity and specificity was developed in DNA extracted from microscope slides, and achieved 100% of efficiency.

Conclusions

DNA extraction by Chelex + NP-40 method from both, cultures of M. tuberculosis and smear slides, resulted in good quantity of interference free DNA, especially in samples with low concentrations of genetic material; therefore, such technique may be used for the molecular diagnosis of tuberculosis.

【 授权许可】

   
2013 de Almeida et al.; licensee BioMed Central Ltd.

【 图 表 】

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