ABSTRACT
Objective: To computationally model the CTX-M-5 ?-lactamase and establish its structure, which is exclusively present in human-associated Salmonella. Methods: The CTX-M-5 aminoacid sequence (Uniprot ID:O65975) of Salmonella enterica subsp. enterica serovar typhimurium was retrieved from UniProt database and subjected to homology modeling using MODELLER 9v7. The homology models were duly validated using RAMPAGE tool by generating Ramachandran plots, ERRAT graphs, and ProSA score. DoGSiteScorer server and ConSurf server were used to detect the cavities, pockets, and clefts to identify conserved amino acid sites in the predicted model. Subsequently, the modeled structure was docked using CLC Drug Discovery Workbench against proven drugs and known inhibitors. Results: Obtained high-quality homology model with 91.7% of the residues in favorable regions in Ramachandran plot and qualified in other quality parameters. Docking studies resulted in a higher dock score for PNK (D-benzylpenicilloic acid) molecule when compared to other reported inhibitors. Conclusion: This in silico study suggests that the compound PNK could be an efficient ligand for CTX-M-5 ?-lactamase and serve as a potent inhibitor of CTX-M-5.
ABSTRACT
BACKGROUND: Increase in extended-spectrum -lactamase(ESBL)-producing Escherichia coli and Klebsiella pneumoniae have been reported in Korea. The aim of this study was to determine the nationwide prevalence of ESBL-producing E. coli and K. pneumoniae, and to investigate the types of ESBLs. METHODS: A total of 2,221 E. coli and 1,128 K. pneumoniae consecutive isolates were yearly collected from 12 hospitals in 1999 and 2000. ESBL production was performed by National Committee for Clinical Laboratory Standards methods and double synergy tests. The type of ESBL was determined by polymerase chain reaction (PCR), isoelectric focusing, and nucleotide sequence analysis. RESULTS: ESBL-producing E. coli and K. pneumoniae isolates were detected from all 12 hospitals participated. The proportion of ESBL-producers was 9.1%(2.0-19.6%) of the E. coli and 29.2% (10.0-60.8%) of the K. pneumoniae isolates. Among the 22 isolates sequenced, SHV-12 was found in six isolates, SHV-2a in three isolates, TEM-52 in five isolates, TEM-106 in three isolates, and each of TEM-15, TEM-20, TEM-43, and TEM-107 in one isolate. CTX-M-14 was also found in one isolate. CONCLUSION: ESBL-producing E. coli and K. pneumoniae are widespred to all levels of Korean hospitals. The most common types of ESBLs in Korea are SHV-12, SHV-2a, and TEM-52. In addition, we also identified new TEM-derived ESBLs.
Subject(s)
Base Sequence , Escherichia coli , Escherichia , Isoelectric Focusing , Klebsiella pneumoniae , Klebsiella , Korea , Pneumonia , Polymerase Chain Reaction , PrevalenceABSTRACT
BACKGROUND: The prevalence of extended-spectrum -lactamase (ESBL)-producing Klebsiella pneumoniae and Escherichia coli has been increased in Korea, but the testing and reporting ESBL-mediated resistance remains unclear. We undertook a study to evaluate the method to screen isolates of ESBL-producing K. pneumoniae and E. coli using cefpodoxime disk. METHODS: Fifty-eight strains of K. pneumoniae and 28 strains of E. coli were tested for production of ESBLs by the double disk synergy test. Susceptibility to cefpodoxime, ceftazidime, cefotaxime, and aztreonam was determined by disk diffusion method. RESULTS: All strains that produced ESBLs were resistant to cefpodoxime, whereas those that not produced ESBLs were susceptible (97%) to this agents. The disk diffusion test exhibited 100% sensitivity and 97% specificity when NCCLS conventional interpretive criteria were used. All other oxyimino- -lactam agents tested were inferior discriminators between the two groups of organisms. When NCCLS ESBL interpretive criteria were used, the disk diffusion test using cefpodoxime exhibited 100% sensitivity and 83% specificity. CONCLUSIONS: Routine disk diffusion susceptibility test with cefpodoxime disk (10g) can be used to detect strains of ESBL-producing K. pneumoniae and E. coli without include supplemental testing for ESBL production.