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Published on: April 25, 2014
Comparison of active and dormant Mycobacterium tuberculosis DosR/DevR regulon polymerase chain reaction-restriction
Faezeh Hamidieh1, Parissa Farnia1, Poopak Farnia1,2
1Mycobacteriology Research Center (MRC), National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Introduction:
This retrospective cross-sectional study aimed to evaluate the differences between the restriction fragment length polymorphism (RFLP) patterns of the dormancy survival regulator (DosR) regulon in latent tuberculosis vs active disease.
Methods:
Sputum samples from 90 patients with active Mycobacterium tuberculosis infection were collected. The presence of the devR, devS, and dosT genes in active and induced dormant M tuberculosis infection was evaluated using polymerase chain reaction (PCR). In addition, the differences between the restriction enzyme digestion of these genes were determined using the RFLP method.
Results:
The devR gene was much more prevalent in dormant than in active samples, with statistical significance set at P = .033. The PCR-RFLP patterns obtained from the effect of the AciI endonuclease on devR, the EaeI and HincII endonucleases on devS, and the HaeIII and AciI endonucleases on dosT showed a statistically significant difference between the active and dormant groups (P = .001, P = .01, P = .008, P = .001, and P = .001, respectively), and this difference was associated with more diverse patterns in the active group.
Discussion:
Results suggested that the DosR regulon may have more nucleotide variations at the active stage. This study is the first to investigate the devR, devS, and dosT genes using PCR-RFLP as a cost-effective and straightforward method.
