Epidemiology and Characteristics of Metallo-beta-Lactamase-Producing Pseudomonas aeruginosa

Metallo-beta-lactamase-producing Pseudomonas aeruginosa (MPPA) is an important nosocomial pathogen that shows resistance to all beta-lactam antibiotics except monobactams. There are various types of metallo-beta-lactamases (MBLs) in carbapenem-resistant P. aeruginosa including Imipenemase (IMP), Verona integron-encoded metallo-beta-lactamase (VIM), Sao Paulo metallo-beta-lactamase (SPM), Germany imipenemase (GIM), New Delhi metallo-beta-lactamase (NDM), Florence imipenemase (FIM). Each MBL gene is located on specific genetic elements including integrons, transposons, plasmids, or on the chromosome, in which they carry genes encoding determinants of resistance to carbapenems and other antibiotics, conferring multidrug resistance to P. aeruginosa. In addition, these genetic elements are transferable to other Gram-negative species, increasing the antimicrobial resistance rate and complicating the treatment of infected patients. Therefore, it is essential to understand the epidemiology, resistance mechanism, and molecular characteristics of MPPA for infection control and prevention of a possible global health crisis. Here, we highlight the characteristics of MPPA.

Detection of Carbapenemases in Clinical Enterobacteriaceae Isolates Using the VITEK AST-N202 Card

BACKGROUND: The rapid and accurate detection of carbapenemase-producing Enterobacteriaceae (CPE) in clinical microbiology laboratories is essential for the treatment and control of infections caused by these microorganisms. This study was performed to evaluate the ability of the VITEK AST-N202 card to detect CPE isolates. MATERIALS AND METHODS: A total of 43 (Klebsiella pneumoniae, n = 37; Escherichia coli, n = 3; and Enterobacter cloacae, n = 3) CPE isolates and 79 carbapenemase-non-producing Enterobacteriaceae (CNE) isolates were included in this study. The CPE isolates harbored KPC-2 (n = 11), KPC-3 (n = 20), GES-5 (n = 5), VIM-2 (n = 2), IMP-1 (n = 1), NDM-1 (n = 2), or OXA-232 (n = 2). Of the 79 CNE isolates, eight K. pneumoniae isolates were resistant to ertapenem, imipenem, and meropenem, while the remaining 71 isolates were susceptible to the carbapenems. Antimicrobial susceptibilities were tested using the VITEK AST-N202 card, and the results were interpreted as positive when the isolates showed resistant or intermediate results. Modified-Hodge tests (MHTs) were performed using ertapenem or meropenem disks for the screening of carbapenemase production. Polymerase chain reaction (PCR) and direct sequencing were used to identify beta-lactamase genes. RESULTS: Sensitivity of MHT with ertapenem and meropenem disks for the detection of carbapenemase was 81.4% (35/43) and 81.4% (35/43), respectively, and a combination with both antibiotic disks increased the sensitivity to 88.4% (38/43). Specificity of the MHT was 100% (79/79) for the CNE isolates. Sensitivity of ertapenem, imipenem, and meropenem as assessed by the VITEK AST-N202 card was 100% (43/43), 93% (40/43), and 95.3% (41/43), respectively. Specificity (89.8%, 71/79) of the test with each carbapenem was improved to 100% (71/71) when eight carbapenem-resistant CNE isolates were excluded from the testing. CONCLUSION: The VITEK AST-N202 card showed high sensitivity for the detection of carbapenemases in Enterobacteriaceae strains. PCR and sequencing experiments for the detection of carbapenemases are recommended when clinical Enterobacteriaceae isolates show non-susceptibility to carbapenems.

Prevalence of the Extended-Spectrum beta-Lactamase and qnr Genes in Clinical Isolates of Escherichia coli / 대한진단검사의학회지

BACKGROUND: This study was performed to investigate the prevalence of qnr genes in clinical isolates of Escherichia coli from Korea that produce extended-spectrum beta-lactamases (ESBLs). METHODS: During the period of May to June 2005, we collected clinical isolates of E. coli that were intermediate or resistant to ceftazidime and/or cefotaxime from 11 Korean hospitals. Antimicrobial susceptibility was determined by the disk diffusion and agar dilution methods. ESBL production was confirmed phenotypically by the double-disk synergy test. ESBL and qnr genes were searched for by PCR amplification, and the PCR products were then subjected to direct sequencing. RESULTS: Double-disk synergy tests were positive in 84.3% (118/140) of ceftazidime- and/or cefotaxime-nonsusceptible E. coli isolates. The most prevalent types of ESBL in E. coli isolates were CTX-M-14 (N=41) and CTX-M-15 (N=58). Other ESBLs were also identified, including CTX-M-3 (N=7), CTX-M-9 (N=8), CTX-M-12 (N=1), CTX-M-57 (N=1), SHV-2a (N=2), SHV-12 (N=17) and TEM-52 (N=4). The qnrA1 and qnrB4 genes were identified in 4 and 7 ESBL-producing isolates, respectively. CONCLUSIONS: CTX-M-type enzymes were the most common type of ESBL in E. coli isolates from Korea, and the qnr genes were not uncommon in ESBL-producing E. coli isolates. Dissemination of E. coli containing both ESBL and qnr genes could compromise the future usefulness of the expanded-spectrum antibiotics for the treatment of infections.

Prevalence of PER-1 Extended-Spectrum beta-Lactamase-Producing Clinical Isolates of Acinetobacter baumannii in a University Hospital, Busan, Korea / 대한임상미생물학회지

BACKGROUND: In recent years, Acinetobacter baumannii isolates acquired resistance to cefepime have increased significantly. The aim of this study was to survey the prevalence of PER-1 extendedspectrum beta -lactamase (ESBL)-producing A. baumannii isolates in a University Hospital, Busan, Korea. METHODS: Antimicrobial susceptibilities were tested by the disk diffusion method, and double disk synergy test was performed for screening of ESBL-production. MICs were determined by agar dilution method. The isoelectric points of beta -lactamases were determined by isoelectric focusing. Transferability of cefepime-resistance were tested by conjugation. blaPER-1 and blaPER-2 alleles were detected by PCR, and the DNA sequences of amplified products were determined by using the dideoxy-chain termination method. RESULTS: Among 51 clinical isolates of A. baumannii intermediate or resistant to cefepime, 10 isolates (19.6%) showed positive results in double disk synergy test. PCR-based experiments detected blaPER-1 gene in all the 10 isolates. All the isolates contained three beta -lactamase bands: pI 5.3, 7.9, and 9.4. MICs of ampicillin, piperacillin, cephalothin, cefoxitin, cefoperazone, ceftazidime, cefotaxime, cefepime, and aztreonam were >256 mg/L, respectively, and them of imipenem were 8-16 mg/L. CONCLUSION: The prevalence of PER-1-producing A. baumannii strains in Busan was less than that of in Seoul. But an outbreak of infection caused by this strain in an intensive care unit shows that spread of PER-1-producing A. baumannii strains can be anticipated in a near future. Prevention of hospital infection by these resistant microorganisms are needed.

Prevalence of CTX-M-type Extended-Spectrum beta-Lactamase-Producing Esherichia coli and Klebsiella pneumoniae Isolates in Korea / 대한임상미생물학회지

BACKGROUND: The aim of this study was to survey the nation wide susceptibilities of Esherichia coli and Klebsiella pneumoniae against cefotaxime and to determine the prevalence of CTX-M-type extended-spectrum beta- lactamases(ESBLs). METHODS: During the period of April to June, 2002, E. coli and K. pneumoniae isolates were collected from 13 hospitals. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production was determined by double disk synergy test. Cefotaxime-resistance of the ESBL-producers was transferred to azide-resistant E. coli J53 by conjugation. MICs of beta- lactam antibiotics were determined by agar dilution method. Searches for blaCTX-M genes were performed by PCR amplication. pIs of beta-lactamases were determined by isoelectric focusing. RESULTS: Ten percents of E. coli and 35 percents of K. pneumoniae isolates among 260 strains of each were intermediate or resistant to cefotaxime. Twenty-three isolates of E. coli and 78 K. pneumoniae isolates showed positive results in the double disk synergy test. One isolate of E. coli and 2 K. pneumoniae isolates harbored blaCTX-M-3 gene, 2 E. coli isolates harbored blaCTX-M-15 gene, and 2 E. coli and 2 K. pneumoniae isolates harbored blaCTX-M-14 gene. CONCLUSION: E. coli and K. pneumoniae isolates producing CTX-M-type ESBLs are not uncommon in Korean hospitals. The spread of CTX-M-type ESBL genes could compromise the future usefulness of 3rd generation cephalosporins and aztreonam for the treatment of E. coli and K. pneumoniae infections.