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Molecular Characterization of Tuf Gene and Antibiotic Susceptibility Patterns of Streptococcus Species in Local Population of Lahore, Pakistan

Author(s): A. Mahmood, A. J. Sami and Q. Ali

Background: Multi Drug Resistance (MDR) is a global problem which is expeditiously increasing. Different antimicrobial drugs have been reported resistant against various Streptococcus species. This has been a threat to local individuals in all over the world specifically in developing countries including Pakistan. Decreased efficiency of drugs due to emergence of new resistance mechanisms in treatment of common infectious diseases like pneumonia and meningitis has been increasing. The cause of the failure of microbial response to standard treatment leading to prolonged illness and high risk of death. This study aims to determine the impact of antibiotics on Streptococcus species by studying the resistance and sensitivity patterns in local population of Lahore. Methods: A community based study has been conducted from different areas of Lahore. A total of 220 clinical samples including oral dental caries, pus, nasopharyngeal, urine and skin has been studied from June 2018 to May 2019. Species identification has been done by microscopic visualization, Gram staining and biochemical identification including catalase, coagulase, bile esculin and NaCl broth test. Antibiotic susceptibility patterns of the clinical isolates have been defined by using commercially prepared antibiotic discs (Streptomycin, Ampicillin, Vancomycin, Oxacillin, Tetracycline, Clindamycin, Chloramphenicol, Gentamicin, Amoxicillin and Azithromycin). Molecular characterization of resistant Streptococcus species has been evaluated by targeting tuf gene. Primers have been designed manually using consensus gene sequence of tuf gene of Streptococcus species from PUBMED and Primer BLAST has been used to analyse the quality of primers. DNA was isolated by phenol Chloroform Method. Polymerase Chain Reaction (PCR) was performed to amplify tuf gene. The amplified product has been evaluated by using Agarose Gel Electrophoresis. Results: Resistance and Sensitivity have been studied against Streptococci according to the mode of the action of different antibiotics. It was observed that resistance of Streptococcus species against the specific drugs is increasing gradually. Exceptionally surprising results have also been observed as some of the species were sensitive to different antibiotics in previously reported studies but currently become resistant. Conclusion: This study concluded Streptococcus species including S. mitis and S. bovis are opportunistic, upon getting the required conditions these become infectious. Malnutrition is one of the major causes of the increasing resistance patterns of Streptococcus species.