Background: Tuberculosis is a critical infectious disease primarily affecting the lungs
and is more common in developing countries. In the 21st century, it forms a significant
problem for world public health especially with the emergence and rising of drug
resistant TB. Microbiological methods are the clue for the laboratory diagnosis. The
ordinary methods for TB identification showed either weak sensitivity as in microscopy
or lateness for many weeks as in culture. The evolution in molecular biology gives a
chance for fast diagnosis of Mycobacterium tuberculosis helping start proper treatment
early and holding its spread. The initial critical step in PCR is DNA extraction.
Objective: The aim to evaluate different extraction methods of Mycobacterium
tuberculosis retrieved directly from sputum samples and from LJ isolates from same
patients and comparing DNA yield using conventional PCR. Methodology: DNA from
32 sputum samples from TB patients extracted by solid, digestion and phenol extraction
methods, DNA from 40 LJ isolates extracted by solid, boiling and Cetyl
trimethylammonium bromide methods. Extracted DNA was evaluated by conventional
PCR. Results: Among 32 sputum samples, the extracted DNA by phenol method was
21/32 (65.62%) with highest DNA yield, digestion method 14/32 (43.75%) and solid
phase method 1/32 (2.5%) with least DNA yield. From 40 MTB LJ culture isolates, the
extracted DNA by boiling method was 28/40 (70%), CTAB method 18/40 (45%) and
solid phase method 2/40 (5%). Conclusion: Phenol method was the best method (mean
rank 2.34) for DNA extraction from sputum samples, while the easy and economic
boiling method was the best method (mean rank 2.45) for DNA extraction from LJ
culture isolates. The worst method of DNA extraction from both sputum and culture was
phase solid method. A greater and easier yield of DNA was obtained from MTB LJ
Culture than sputum.