Evaluation of Simple and Cost-Effective DNA Preparation and Subsequent PCR Amplification for Clinically Relevant Mycobacteria
Journal Title: Journal of Advances in Medicine and Medical Research - Year 2015, Vol 8, Issue 2
Abstract
Aim: Slow growth rate in culture renders the traditional isolation, identification, and drug susceptibility testing of clinically important mycobacteria inadequate when there is an urgent need for a precise diagnosis in order to initiate patient treatment. Molecular methods all rely on mycobacterial DNA isolation which in turn has become an essential step of the process. Our study aimed to evaluate DNA isolation protocols from mycobacteria of clinical interest. Methods: Therefore, in order to determine an optimal method we evaluated 8 inexpensive, rapid and easy DNA isolation methods from 30 mycobacterial cultures (10 Mycobacterium tuberculosis and 20 Non-tuberculous Mycobacteria) for subsequent direct detection by PCR. Results: Six of those 8 methods reliably allow the isolation of good DNA yields and quality, the optimal protocol being the one that includes a 1% Triton X-100 lysis solution. Protocols using SDS 1% as a lysis solution did not yield DNA suitable for PCR amplification. Conclusion: Six of the methods we evaluated can easily be implemented in resource limited settings for routine use, potentially contributing to a better management of mycobacterial infections.
Authors and Affiliations
Veronica J. Gómez, Angela M. Guzman, Gloria I. Mejia, Diego H. Caceres, Jaime A. Robledo, Francois Rouzaud
Keywords
Related Articles
- EP ID EP347657
- DOI 10.9734/BJMMR/2015/16945
- Views 68
- Downloads 0
Role of Tumor Markers in Oral Cancer: An Overview
The oral mucosa represents the first part of the digestive tract and is exposed to various exogenous toxins. Exposure for longer duration can lead to changes that lead to potentially malignant diseases or cancers / Tumor...
Patient Satisfaction with Treatment Method: A Determinant of the Efficacy of Cognitive Behaviour Therapy Compared with Psycho-pharmacological Intervention Therapy
The first aim of the research was to compare the Cognitive Behavior Therapy (CBT) with psychopharmacological intervention (PI) and to find out its effectiveness on Iranian women with Anxiety disorders (general anxiety di...
Comparison of the Quality of Life of Vulnerable Children Resident in Household and Those Resident in Institution in Jos Nigeria
Background: There are over 143 million orphans globally, however, sub-Saharan Africa/Asian regions account for over 80% of the global burden (143 million orphans), Nigeria inclusive with over 10 million orphans. This has...
Epidemiology of Cardiac Arrests in Airports: Four Years Results of the French National Cardiac Arrest Registry
Aims: To describe and analyse cardiac arrests occurring in airports, all chain of survival steps and their survival including a neurological outcome assessment at day 30 and to compare these results between airports staf...
Prevalence of Hashimoto’s Thyroiditis and Its Association with Vitamin D Deficiency in West Bengal, India
Objective: The aim of our study is to screen the prevalence of Hashimoto’s Thyroiditis and to evaluate a correlation between 25(OH)Vitamin D deficiency and AITD in hypothyroid patients of West Bengal. Study Design: Ethic...