TEM & SHV genes in extended spectrum beta-lactamase producing Klebsiella species beta their antimicrobial resistance pattern.

ABSTRACT

BACKGROUND & OBJECTIVES: Extended spectrum beta-lactamases (ESBLs) are often plasmid mediated derived from mutations in the classic TEM and SHV genes by one or more amino acid substitution around the active site. Detection of TEM and SHV genes by molecular methods in ESBL producing bacteria and their pattern of antimicrobial resistance can provide useful information about its epidemiology and risk factors associated with these infections. We investigated the presence of TEM and SHV genes in ESBL producing Klebsiella spp. and their antimicrobial resistance pattern in cases of neonatal septicaemia in a tertiary care hospital. METHODS: A total of 130 clinical isolates of Klebsiella spp. isolated from septicaemic neonates of a neonatal intensive care unit (NICU) from a tertiary care hospital in north India, were screened for ESBL production by combined disk diffusion method. PCR was used to detect TEM and SHV genes in ESBL positive isolates. Isoelectric points of ESBL enzymes from a few isolates (n = 6) were noted for typing of ESBL by isoelectric focusing. RESULTS: Of the 64 ESBL producing Klebsiella spp. isolates, 17 (26.5%) had both TEM and SHV genes, 31 (48.4%) had TEM alone and 13 (20.3%) had SHV gene alone. Three (4.6%) ESBL positive isolates were negative for both TEM and SHV. Isolates with both TEM and SHV genes were highly resistant to antibiotics used. Degree of resistance for 3(rd) generation cephalosporins was also high in these isolates. Six randomly selected isolates were subjected to isoelectric focussing. Results of isoelectric focussing were comparable with PCR. INTERPRETATION & CONCLUSION: Presence of TEM gene in ESBL producing Klebsiella spp. was more common than SHV gene. Frequency of antibiotic resistance was high in isolates having both TEM and SHV genes.