Diversity of SHV and TEM beta-lactamases in Klebsiella pneumoniae: Gene evolution in northern Taiwan and two novel beta-lactamases, SHV-25 and SHV-26

A total of 113 blood culture isolates of Klebsiella pneumoniae from 10 hosp itals in northern Taiwan were studied for SHV and TEM beta -lactamase produ ction. bla(SHV) was amplified from all isolates by PCR. TEM-type resistance , was found in 32 of the isolates and was of the TEM-I type in all isolates . SHV-1, -2, -5, -11, and -12 and two novel enzymes were identified. These novel enzymes were designated SHV-25 and SHV-26 and had pis of 7.5 and 7.6, respectively. Amino acid differences in comparison to the amino acid seque nce of bla(SHV-1) were found at positions T18A (ThrACC --> AlaGCC), L35Q (L euCTA --> GluCAA), and M129V (MetATG --> ValGTG) for SHIV-25 and at positio n A187T (AlaGCC --> ThrACC) for SHV-26. The results of substrate profiles a nd MIC determinations showed that the novel enzymes did not hydrolyze exten ded-spectrum cephalosporins, rendering the isolates susceptible to these ag ents. Inhibition profiles revealed that the 50% inhibitory concentration fo r SHV-26 was higher than those for SHV-1 and SHV-25, resulting in an interm ediate resistance to amoxicillin-clavulanic acid. Forty-nine ribotypes were identified, suggesting that major clonal spread had not occurred in any of the hospitals. According to the amino acid sequence, SHV beta -lactamases in Taiwan may basically be derived through stepwise mutation from SHV-1 or SHV-11 and further subdivided by four routes. The stepwise mutations initia ted from SHV-1 or SHV-11 to SHV-2, SHV-5, and SHV-12 comprise the evolution ary change responsible for extended-spectrum beta -lactamase (ESBL) product ion in Taiwan. The stepwise mutations that lead to a non-ESBL (SHV-25) and the beta -lactamase (SHV-26) with reduced susceptibility to clavulanic acid are possibly derived from SHV-11 and SHV-1, respectively. The results sugg est a stepwise evolution of SHV beta -lactamases in Taiwan.