Session:

Antibiotics

Abstract No.:

56.055

Title:

Genotypic Distribution of Plasmid Encoded Extended Spectrum Beta-lactamases from Lahore, Pakistan

Author(s):

S. Riaz; department of microbiology and molecular genetics, department of microbiology and molecular genetics, Lahore/PK

Abstract:

Background: Successful spread of ESBL can be attributed to the fact that genes encoding for ESBL are often located on self-transmissible or mobilizable broad range plasmids. There is great diversity in type of gene producing beta-lactamase, such as TEM, SHV, OXA, PER, VEB, BES, CTX-M and others.  Here genotypic distribution of plasmid encoded extended spectrum Beta-lactamases from Lahore, Pakistan were studied.
Methods: DNA Amplification for TEM, SHV and OXA-1 encoding genes was performed using singlplex and multiplex PCR, plasmid isolation, horizontal gene transfer, isoelectric focuing and amino acids were calculated in local isolates of E.coli and Klebiella spp.
Results: In 260 clinical isolates of E. coli suspected to be ESBL producers OXA (19.2%) and TEM/OXA (44.2%) were more prevalent. Of the 40 clinical isolates of Klebsiella spp 92.5% demonstrated SHV-specific products. In 87 environmental isolates of E. coli 58.6% TEM/OXA and out of 13 environmental isolates of Klebsiella spp exhibited 76.92% SHV. All tested isolates showed high rate of resistance transfer though horizontal gene transfer mechanism. Isolectric focusing was performed that give isoelectric points for specific beta-lactamase.
Conclusion: ESBL plasmid encoded genes were easily transfered through horizontal gene transfer mechanism.  The best results were obtained from PCR and DNA sequencing as compared to calculating isolectric points (pI). If somehow multiplex PCR of ESBL will applied in local diagnostic labs this helps in early detection and phenotypic antibiotic therapy against this infection.