Cultural and molecular characterization of tetracycline resistant microflora associated with dental caries

Luhana K.K.¹*, Patel H.D.²*
¹Shree M. and N. Virani Science College, Rajkot, Gujarat
²Disha life Sciences Pvt. Ltd, 301, Trade Center, Gurukul Road, Ahemedabad
* Corresponding Author : hp3948@gmail.com

Received : -     Accepted : -     Published : 15-06-2010
Volume : 2     Issue : 1       Pages : 1 - 7
Genetics 2.1 (2010):1-7
DOI : http://dx.doi.org/10.9735/0975-2862.2.1.1-7

Keywords : Dental caries, tet K gene, Bacteria, FAME analysis, Tetracycline, PCR, RFLP
Conflict of Interest : None declared

Dental caries is the most common chronic disease. Tetracycline is commonly used in dental practice as a prophylactic agent and for treatment of oral infections. The wide use of tetracycline had resulted in a major increase in the rate of tetracycline resistance among bacteria. Substantial epidemiologic evidence links Streptococcus mutans to caries; the pathobiology of caries may involve more complex communities of bacterial species. Molecular methods for bacterial identification and enumeration now make it possible to more precisely study the micro biota associated with dental caries. The purpose of this study was to characterize the tetracycline resistant microorganism associated with the dental caries using cultural & molecular method. The first step involved 20 samples from patients with different level of dental caries. From which 33 different isolates were obtained, out of which 12 showed resistances to tetracycline and 5 were having maximum MIC indicating that they are highly resistant. These resistant microorganisms were identified to the species level by growing them on a suitable medium using the antibiotic tetracycline and were analyzed with the help of FAME analysis, which indicated that different strains of the Pseudomonas aeruginosa and Bacillus subtilis were present.The second step involved identifying the tet k gene in the cultures. Tet k was amplified under standard conditions by using specific primers results. Interestingly, the results showed that tet k gene was present in all the sample. Tet k gene is involved in imparting the resistant to the tetracycline by acting as a efflux pump. RFLP analyses were used in order to identify the polymorphic forms of the tet k. The HindIII digest suggest that high resistance to tetracycline was acquired due to some mutations in the genome.