IDENTIFICATION OF BACTERIA CAUSING NECROTIC PULP WITH 16S rRNA GENE POLYMERASE CHAIN REACTION AND ANTIBIOTIC RESISTANCE TESTING AT THE DENTAL HOSPITAL IN SEKELOA, BANDUNG, INDONESIA
Keywords:Identification of bacteria, Necrotic pulp, Polymerase chain reaction of 16S rRNA gene, Sequencing, Antibiotic resistance test
Objective: This study aims to identify the bacteria that cause necrotic pulp in teeth of dental patients and test resistance of the bacteria found to antibiotics.
Methods: Bacteria were taken with paper points that were inserted into the root canal of patients at the Dental Hospital of the Faculty of Dentistry, Universitas Padjadjaran, Sekeloa, Bandung. Bacteria were cultured and Gram-stained. Bacterial DNA was isolated to be identified by polymerase chain reactions 16S rRNA method against known sequences of bacterial DNA. Then, identified bacteria were tested for antibiotic resistance to tetracycline (30 Î¼g), clindamycin (2 Î¼g), amoxicillin (10 Î¼g), and ampicillin (10 Î¼g).
Results: The 16s rRNA gene fragment of the main bacterium found was in 98% homology with 16S rRNA gene database in http://blast.ncbi.nlm. nih.gov, i.e., Pseudomonas aeruginosa and Acinetobacter schindleri. The results from inhibition zone of each antibiotic were 20.12 mm, 8.97 mm, 8.12 mm, and 8.03 mm for tetracycline, clindamycin, amoxicillin, and ampicillin, respectively, to P. aeruginosa. While inhibition zone of tetracycline to A. schindleri was 37.7 mm.
Conclusion: Based on the study results, P. aeruginosa from patients with necrotic pulp samples were resistant to clindamycin, amoxicillin, ampicillin, and decreased activity to tetracycline. While Acinetobacter schindleri was still sensitive to tetracycline.
Garg N, Garg A. Textbook of Endodontics. 2nd ed. New Delhi: Brothers Medical Publishers; 2011
Pradono J, Soemantri S. Survei kesehatan nasional: Survei kesehatan rumah tangga (SKRT) 2004. Volume 3. Jakarta: Departemen Kesehatan Republik Indonesia.
Rocassa IN, Siqueira JF Jr. Root Canal Microbiota of Teeth with Chronic Apikal Periodontitis. J Clin Mikrobial 2008;46(11): 3599-606.
Rhodes JS. Advanced Endodontics. 1st ed. London and New York: Taylor & Francis Publishers; 2006.
Jaju S, Jaju PP. Newer Root Canal Irrigants in Horizon : A Review. Intl J Dent 2011;2011:851359.
Mullany P, Allan E, Warburton PJ. Tetracycline resistance genes and mobile genetic elements from the oral metagenome. Clin Microbiol Infect 2012; 18 (4): 58â€“61.
Kementerian Kesehatan Indonesia. Profil kesehatan Indonesia tahun 2010. Jakarta: Kementerian Kesehatan Republik IndonesiaI; 2011.
Balcht AL, Smith RP, editors. Pseudomonas aeruginosa: Infections and Treatment. New York: Marcel Dekker; 1994.
Dortet L, Legrand P, Soussy CJ, Cattoir V. Bacterial identification, clinical significance, and antimicrobial susceptibilities of Acinetobacter ursingii and Acinetobacter schindleri, two frequently misidentified opportunistic pathogens. J Clin Microbiol 2006;44(12); 4471-4478.
Siqueira JF. Treatments of Endodontics Infections. 1st ed. Berlin: Quintessence Publishing; 2011.
Coggan KA, Wolfgang MC. Global regulatory pathways and cross-talk control Pseudomonas aeruginosa environmental lifestyle and virulence phenotype. Curr Issues Mol Biol 2012; 14(2):47-70.
Riou M, Carbonnelle S, Avrain L, Mesaros N, Pirnay JP, Bilocq F, et al. In vivo development of antimicrobial resistance in Pseudomonas aeruginosa strains isolated from the lower respiratory tract of intensive care unit patients with nosocomial pneumonia and receiving antipseudomonal therapy. Int J Antimicrob Agents 2010; 36(6):513-22.
Brown SP, Cornfort DM, Mideo N. Evolution of virulence in opportunistic pathogens: generalism, plasticity and control. Trends Microbiol 2012; 20(7):336-42.
Skucaite, N, Peciuliene V, Vitkauskiene A, Machiulskiene V. Susceptibility of Endodontic Pathogens to Antibiotics in Patients with Symptomatic Apical Periodontitis. JOE 2010; doi:10.1016/j.joen.2010.04.009
Garner JS, Jarvis WR, Emori TG, Horan TC, Hughes JM. Definition for nosocomial infections. Am J Infect Control 1988;16:128â€“40.
Aloush V, Navon-Venezia S, Seigmen-Igra Y, Cabili S, Carmeli Y. Multidrug resistant Pseudomonas aeruginosa: risk factors and clinical impact. Antimicrob Agents Chemother 2006; 50(1):43-8.
Bredenstein EBM, de la Fuente-Nunez C, Hancock REW. Pseudomonas aeruginosa: all roads lead to resistance. Trends Microbiol 2011; 19(8):419-25.
Chapter in book : Meletis G, Bagkeri M. Infection Control.In : Basak S, editor. Pseudomonas aeruginosa: Multi-Drug-Resistance Development and Treatment Options. InTech publisher 2013:33-56. Available from: http://www.intechopen.com/books/infection-control/pseudomonas-aeruginosa-multi-drug-resistance-development-and-treatment-options.
Clinical and Laboratory Standards Institute. 2006. Performance standards for antimicrobial susceptibility testing; sixteenth informational supplement, vol. 26, no. 3. M100-S16. Clinical and Laboratory Standards Institute, Wayne, PA.1988;16(3):128-40.
Aloush V, Navon-Venezia S, Seigman-Igra Y, Cabili S, Carmeli Y. Multidrug-resistant Pseudomonas aeruginosa: Risk factors and clinical impact. Antimicrob Agents Chemother 2006;50(1):43-8.
Bredenstein EB, de la Fuente-Nunez C, Hancock RE. Pseudomonas aeruginosa: All roads lead to resistance. Trends Microbiol 2011;19(8):419-25.
Meletis G, Bagkeri M. Pseudomonas aeruginosa: Multi-drug-resistance development and treatment options. In: Basak S, editor. Infection Control. Croatia: InTech Publisher; 2013. p. 33-56.
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