Detection of Pel A Gene in P. aeruginosa from Clinical Samples Using Polymerase Chain Reaction with Reference to Biofilm Production In N.E India.
Journal Title: PARIPEX-Indian Journal of Research - Year 2015, Vol 4, Issue 10
Abstract
Objectives: The aims of the current study were to determine the biofilm formation in P. aeruginosa clinical isolates and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in N.E India. Methods: A total of 45 clinical isolates were isolated from Silchar medical college and hospital, Silchar, Assam followed by antibiotic susceptibility test for all the strong, intermediate, weak and non-biofilm producing strains against aminoglycosides and beta lactam antibiotics. Using biofilm assays, the ability of P. aeruginosa isolates to form biofilm was estimated and according to the OD values, isolates was classified into weak, moderate or strong biofilm producers. A PCR assay was carried out for detection of PelA gene which was involved in formation of biofilm among the clinical isolates. Results: Of the 47 isolates, 42(89.3%) isolates confirmed the production of biofilm amongst which 34 (72.34%) were strong, 5 (10.63%) intermediate and 3(7.14%) weak biofilm producers. The aminoglycosides set of antibiotics represented 78.5% resistant towards gentamycin, 83.3 % Tobramycin, 80.9% Amikacin and finally 73.8 % towards Kanamycin. The beta lactam antibiotics exhibited the resistant pattern higher in biofilm producing strains where ceftazidime expressed 69.0% resistance, cefepime 61.9%, cefixime 66.6% and piperacilin 57.1%. Further, in the present study PelA gene was expressed heavily (80%) amongst the biofilm producing isolates and those associated with the polysaccharide stage of biofilm development and maintenance (97%). It was also observed that the phenotypically positive isolates for production of biofilms in vitro were genotypically positive for expression of PelA gene while those isolates phenotypically producing biofilm and harbouring PelA gene were also found to be resistant towards aminoglycosides and beta lactam antibiotics. Conclusions: The present study showed that most of the biofilm producing strains were highly resistant to even higher generations of aminoglycosides and beta lactam group of antibiotics, therefore, more work is needed to fully elucidate this antibiotic resistance mechanism in biofilms and develop new therapeutic strategies.
Authors and Affiliations
Indu Sharma, Daijee Choudhury
Keywords
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