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Free Content Determination of polymerase chain reaction efficiency for diagnosis of tuberculous meningitis in Chelex-100® extracted DNA samples

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Abstract:

SETTING: Polymerase chain reaction (PCR) offers great promise for the rapid, sensitive and specific diagnosis of tuberculous meningitis (TBM). However, the isolation of DNA of high quantity and quality from cerebrospinal fluid (CSF) samples is critical for successful PCR assays.

OBJECTIVE: To develop and use a single-tube method for the isolation of PCR-compatible DNA from Mycobacterium tuberculosis using Chelex-100® chelating resin, which does not require organic solvents or detergents.

DESIGN: The study focused on the standardisation of a suitable Chelex protocol and its evaluation in 32 CSF samples from TBM and non-TBM subjects. A simultaneous comparison was made with the conventional phenol/chloroform extraction method.

RESULT: PCR was found to be more sensitive, more rapid and less technically demanding with the Chelex protocol than the conventional phenol/chloroform extraction method (sensitivity 84.2% vs. 73.6%).

CONCLUSION: The single-tube method and the simplicity of the procedure permits early and reliable diagnosis of TBM and makes it an attractive method for routine laboratory assays.

Keywords: Chelex-100; DNA extraction; PCR; TBM; diagnosis

Document Type: Regular Paper

Affiliations: 1: Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur, India 2: Environmental Genomics Unit, National Environmental Engineering Research Institute, Nehru Marg, Nagpur, India

Publication date: 2010-08-01

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  • The International Journal of Tuberculosis and Lung Disease publishes articles on all aspects of lung health, including public health-related issues such as training programmes, cost-benefit analysis, legislation, epidemiology, intervention studies and health systems research. The IJTLD is dedicated to the continuing education of physicians and health personnel and the dissemination of information on tuberculosis and lung health world-wide.

    Certain IJTLD articles are selected for translation into French, Spanish, Chinese or Russian. They are available on the Union website

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