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Original Article

AJVS. 2014; 43(1): 82-89


Comparison of Different DNA Extraction Protocols for Molecular Diagnosis of Mycobacterium Tuberculosis

Sara M. El-Shannat, Samy A. Khalil.




Abstract

Tuberculosis remains a global epidemic, especially in developing countries; early diagnosis plays a critical role in controlling the disease. Traditional method is not reliable because isolation and identification of mycobacteria may take up to several weeks or more in achieving results. The aim of this study is to compare the efficacy of different protocols of mycobacterial DNA extraction in order to standardize a trustable DNA extraction protocol providing high quality DNA for molecular diagnosis. DNA was extracted using six different protocols. PCR was performed using primer of IS6110 that specific for all the members of M. tuberculosis complex. The highest DNA extraction efficiency (68.75%) was observed using the protocol number 3, and DNA extraction was proved to give a higher accuracy to IS6110 PCR in comparison with the other protocols. It could be concluded that most of DNA extraction kit dependent protocols are costly but of high quality DNA extraction, otherwise some of the classical methods are easy, low cost and simple but the purity of the DNA of low quality.

Key words: Mycobacterium tuberculosis; Polymerase Chain Reaction (PCR); IS6110.






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