Klebsiella pneumoniae ST147: and then there were three carbapenemases.

Gram-negative bacteria containing three different carbapenemases are extremely rare. Klebsiella pneumoniae (N22-925) with KPC-2, NDM-1, and OXA-48 was obtained from a Canadian patient with recent hospitalization in Romania. Short and long read whole genome sequencing showed that the blaKPC-2 was situated on a 214 kb IncFIB(K)/IncFII(K) plasmid, the blaNDM-1 on a 104 kb IncFIB (pQil)/IncFII(K) plasmid, and the blaOXA-48 on a 64 kb IncL plasmid. These plasmids were conjugated to Escherichia coli J53. N22-925 belonged to a unique ST147 cluster that is likely endemic in Romania. This case emphasizes the need for rapid carbapenemase screening in patients from endemic regions. We described the first complete genome sequence of a K. pneumoniae isolate with three different carbapenemases, providing a reference for future studies on this rarely reported occurrence.

Molecular epidemiology of Escherichia coli causing bloodstream infections in a centralized Canadian region: a population-based surveillance study.

OBJECTIVES: Escherichia coli is a leading cause of bloodstream infections worldwide, and is responsible for substantial patient morbidity, mortality and healthcare expenditure. Understanding the molecular epidemiology of E. coli will aid in designing superior treatment and prevention strategies. METHODS: We undertook a population-based surveillance study describing the clinical factors, susceptibility patterns, incidence rates and geographical distribution of sequence types (STs) among E. coli isolates (n = 686) causing incident bloodstream infections in a centralized Canadian region during 2016. STs were identified using a seven-single-nucleotide-polymorphism quantitative PCR (n = 422) and sequencing of certain house-keeping genes (n = 249). RESULTS: The annual population incidence rate of E. coli bloodstream infections was 48.8/100 000 patient years, and five dominant clones (ST131, ST73, ST69, ST95 and ST1193) accounting for 55% (378/686) of the population were identified, each with a specific geographical distribution within Calgary. ST131 was the most common (overall incidence rate of 10.4/100 000 patient years), an antimicrobial-resistant (AMR) clone affecting mainly the elderly and the very young. ST131 was common among residents in long-term care with an incidence rate of 312.5/100 000 patient years. ST73 was associated with community infections in the elderly, while ST69 and ST95 had increased incidence rates among females. ST1193 was the second most AMR clone and was associated with bloodstream infections in elderly males. CONCLUSIONS: This study showed that E. coli clones have unique characteristics in a well-defined human population. The elimination of ST131 would substantially decrease the overall incidence rate and AMR burden among E. coli bloodstream infections in the Calgary region, leading to considerable public health benefits.

Klebsiella pneumoniae ST307 with OXA-181: threat of a high-risk clone and promiscuous plasmid in a resource-constrained healthcare setting.

INTRODUCTION: Klebsiella pneumoniae with OXA-48-like enzymes were introduced into Tshwane Tertiary Hospital (TTH) (Pretoria, South Africa) during September 2015, causing nosocomial outbreaks. METHODS: PCR methodologies and WGS were used to characterize K. pneumoniae with carbapenemases (n = 124) from TTH (July 2015-December 2016). RESULTS: PCR was used to track K. pneumoniae ST307 with OXA-181 among 60% of carbapenemase-positive isolates in different wards/units over time and showed the transmission of IncX3 plasmids to other K. pneumoniae clones. WGS identified different ST307 clades: 307_OXA181 (consisting of two lineages, A and B) with OXA-181 on IncX3 plasmids (named p72_X3_OXA181) and clade 307_VIM with VIM-1 on IncFII plasmids. Clade 307_OXA181 lineage B was responsible for the rapid increase and transmission of OXA-181 K. pneumoniae in various wards/units throughout TTH, while the numbers of clade 307_OXA181 lineage A and clade 307_VIM remained low. Separate outbreaks were due to K. pneumoniae ST17 and ST29 with p72_X3_OXA181 plasmids. CONCLUSIONS: The high-risk clone K. pneumoniae ST307 with OXA-181 rapidly spread to different wards/units despite infection and prevention measures. ST307 clades and lineages seemingly acted differently in outbreak situations. This study also highlighted the threat of promiscuous plasmids such as p72_X3_OXA181.

Deciphering risk factors for blood stream infections, bacteria species and antimicrobial resistance profiles among children under five years of age in North-Western Tanzania: a multicentre study in a cascade of referral health care system.

BACKGROUND: Blood stream infections (BSIs) cause a complex cascade of inflammatory events, resulting in significant morbidity and mortality in children in Tanzania. This study was designed to delineate circulating bacterial species, antimicrobial resistance (AMR) profiles and risk factors for BSIs and mortality among children in the cascade of referral health care facilities so as to guide comprehensive BSIs management. METHODS: A multiple cross sectional analytical study was conducted between July 20, 2016 to October 04, 2017 involving 950 children less than five years of age in the North-western part of Tanzania. Children with clinical features suggestive of BSIs were included. Demographic, clinical and laboratory information on culture and antimicrobial susceptibility testing was collected from children; and analyzed using STATA version 13.0 software. RESULTS: The prevalence of BSIs among children was 14.2% (95% CI: 12.1-16.6%), with specific prevalence in the district, regional and tertiary hospitals being 8.3, 6.4 and 20.0%, respectively. The most common bacterial pathogens isolated from 135 culture-positive children were Klebsiella pneumoniae (55, 40.4%), Staphylococcus aureus (23, 17.0%), and Escherichia coli (17, 12.6%). Multi-drug resistance (MDR) was higher in isolates from children at Bugando Medical Centre (BMC) tertiary hospital than isolates from district and regional hospitals [OR (95% CI): 6.36 (2.15-18.76); p = 0.001]. Independent risk factors for BSIs were neonatal period [OR (95% CI): 1.93 (1.07-3.48); p = 0.003] and admission at BMC [2.01 (1.08-3.74); p = 0.028)]. Approximately 6.6% (61/932) of children died, and risk factors for mortality were found to be children attending BMC [OR (95% CI): 4.95 (1.95-12.5); p = 0.001)], neonatal period [OR (95% CI): 2.25 (1.02-5.00); p = 0.045)], and children who had blood culture positive results [OR (95% CI): 1.95 (1.07-3.56); p = 0.028)]. CONCLUSIONS: The prevalence of BSIs (14.2%) in this multi-centre study is high and predominantly caused by the MDR K. pneumoniae. Priority interventional measures to combat BSIs and mortality, specifically among neonates at BMC are urgently recommended.

Multicentre evaluation of significant bacteriuria among pregnant women in the cascade of referral healthcare system in North-western Tanzania: Bacterial pathogens, antimicrobial resistance profiles and predictors.

OBJECTIVES: The aim of this multicentre study was to evaluate the magnitude of significant bacteriuria (SB) as well as the implicated bacterial pathogens, antimicrobial resistance (AMR) profiles and risk factors for SB among pregnant women attending different levels of healthcare facilities (HCFs) in Tanzania in order to guide antimicrobial therapy and preventive measures. METHODS: Information on sociodemographic and clinical characteristics, midstream urine culture and antimicrobial susceptibility testing was collected from 1828 pregnant women between March 2016 and May 2017. Data were analysed using STATA v.13.0 software. RESULTS: The prevalence of SB among pregnant women was 17.7% (323/1828; 95% CI 16.0-19.5%), with a predominance of Escherichia coli (164/323; 50.8%), Klebsiella spp. (55/323; 17.0%) and Staphylococcus aureus (28/323; 8.7%). Moreover, 37.5% (121/323) of bacteria were multidrug-resistant [84.3% (102/121) Gram-negative bacteria and 15.7% (19/121) in Gram-positive bacteria; P<0.001]. Third-generation cephalosporin resistance in E. coli, Klebsiella spp. and other Enterobacteriaceae was 13.4%, 21.8% and 27.5%, respectively, and was higher in strains from a tertiary hospital (OR=3.27, 95% CI 1.02-10.49; P=0.046) compared with lower HCFs. Predictors of SB among pregnant women were lack of formal occupation, current hospital admission and presence of co-morbidities. CONCLUSIONS: The prevalence of SB among pregnant women in this study was high (17.7%) and was within the same range reported 10 years ago in a single-centre baseline study. However, there is an increase in AMR in the cascade of referral healthcare system, underscoring the need for health facility level-specific antimicrobial stewardship.

Incidence, risk factors, and outcomes for Enterococcus spp. blood stream infections: a population-based study.

BACKGROUND: Enterococci are a clinically significant cause of bloodstream infections (BSI), particularly in the nosocomial setting. The purpose of this study was to characterize the incidence, risk factors for acquisition, microbiological characteristics and mortality of enterococcal BSI within the well-defined population of a large Canadian health region. METHODS: Surveillance for all episodes of enterococcal BSI occurring among residents of the Calgary Health Zone (population 1.2 million) between 2000 and 2008 was conducted using an electronic surveillance system. Clinical features, microbiology, and outcomes were obtained. RESULTS: A total of 710 incident episodes of enterococcal BSI were identified for an annual incidence of 6.9 episodes per 100,000; the incidences of E. faecalis and E. faecium BSI were 4.5, and 1.6 per 100,000, respectively. Enterococcus faecalis infections were associated with a urinary focus, genitourinary malignancy, and abnormal genitourinary anatomy. E. faecium infections were associated with a gastrointestinal focus. Resistance to ampicillin, vancomycin and ciprofloxacin was higher in E. faecium infection. The overall case fatality rate was 22.8%, and was higher for E. faecium infection. CONCLUSIONS: This is the second population-based study to assess the risk factors for enterococcal BSI and compare the characteristics of infection with E. faecalis and E. faecium. Results suggest that BSI with E. faecalis and E. faecium should be regarded as two clinically different entities with unique sets of risk factors and microbiologic characteristics.

Population-based assessment of the incidence, risk factors, and outcomes of anaerobic bloodstream infections.

BACKGROUND: Anaerobes are a relatively uncommon but important cause of bloodstream infection. However, their epidemiology has not been well defined in non-selected populations. We sought to describe the incidence of, risk factors for, and outcomes associated with anaerobic bacteremia. METHODS: Population-based surveillance for bacteremia with anaerobic microorganisms was conducted in the Calgary area (population 1.2 million) during the period from 2000 to 2008. RESULTS: A total of 904 incident cases were identified, for an overall population incidence of 8.7 per 100,000 per year; 231 (26 %) were nosocomial, 300 (33 %) were healthcare-associated community-onset, and 373 (41 %) were community-acquired. Elderly males were at the greatest risk. The most common pathogens identified were: Bacteroides fragilis group (3.6 per 100,000), Clostridium (non-perfringens) spp. (1.1 per 100,000), Peptostreptococcus spp. (0.9 per 100,000), and Clostridium perfringens (0.7 per 100,000). Non-susceptibility to metronidazole was 2 %, to clindamycin 17 %, and to penicillin 42 %. Relative to the general population, risk factors for anaerobic bloodstream infection included: male sex, increasing age, a prior diagnosis of cancer, chronic liver disease, heart disease, diabetes mellitus, stroke, inflammatory bowel disease, human immunodeficiency virus (HIV) infection, chronic obstructive pulmonary disease (COPD), and/or hemodialysis-dependent chronic renal failure (HDCRF). The 30-day mortality was 20 %. Increasing age, nosocomial acquisition, presence of malignancy, and several other co-morbid illnesses were independently associated with an increased risk of death. CONCLUSION: Anaerobic bloodstream infection is responsible for a significant burden of disease in general populations. The data herein establish the extent to which anaerobes contribute to morbidity and subsequent mortality. This information is key in developing preventative, empiric treatment and research priorities.

Metallo-ß-lactamase-producing Pseudomonas aeruginosa in the Netherlands: the nationwide emergence of a single sequence type.

Recently, the first outbreak of clonally related VIM-2 metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa in a Dutch tertiary-care centre was described. Subsequently, a nationwide surveillance study was performed in 2010-2011, which identified the presence of VIM-2 MBL-producing P. aeruginosa in 11 different hospitals. Genotyping by multiple-locus variable-number tandem-repeat analysis (MLVA) showed that the majority of the 82 MBL-producing isolates found belonged to a single MLVA type (n = 70, 85%), identified as ST111 by multilocus sequence typing (MLST). As MBL-producing isolates cause serious infections that are difficult to treat, the presence of clonally related isolates in various hospitals throughout the Netherlands is of nationwide concern.

Molecular characteristics of extended-spectrum ß-lactamase-producing Escherichia coli from Rio de Janeiro, Brazil.

Twenty-five extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli clinical isolates from Rio de Janeiro, Brazil were characterized by isoelectric focusing, PCR and sequencing of bla(ESBL) genes, plasmid-mediated quinolone resistance determinants, phylogenetic groups, replicon typing, pulsed-field electrophoresis, and multilocus sequencing typing. Twenty-three (92%) ESBL-producing E. coli isolates were positive for bla(CTX-M) genes, aac(6')-Ib-cr, and qnrB. Genetic relatedness of ESBL producers clustered seven (28%) CTX-M-15-producing isolates as sequence type (ST)410, clonal complex (CC) 23, and two (8%) as clone O25-ST131. Our results illustrate the predominance of phylogroup A (52%), ST410 (CC 23) and CTX-M-15 among ESBL-producing E. coli isolates from hospitals in Rio de Janeiro.

First outbreak of VIM-2 metallo-ß-lactamase-producing Pseudomonas aeruginosa in The Netherlands: microbiology, epidemiology and clinical outcomes.

This study was designed to investigate the prevalence and characteristics of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa in a tertiary care centre in The Netherlands, a country that is considered to have a low prevalence of antibiotic-resistant bacteria. Imipenem-resistant P. aeruginosa isolates cultured from clinical specimens during 2008-2009 were analysed phenotypically and molecularly by polymerase chain reaction (PCR) with sequencing. Genotyping was performed by multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA). Clinical information was obtained by electronic chart review for all patients infected or colonised with an imipenem-resistant P. aeruginosa isolate that was included in the study. In total, 106 imipenem-resistant P. aeruginosa isolates were included. The bla(VIM-2) gene was detected in 35/106 isolates (33%) and was associated with integrons. Compared with non-MBL-producing imipenem-resistant P. aeruginosa, VIM-2 MBL-producing isolates showed higher rates of multidrug resistance. Patients with VIM-2 MBL-producing isolates were more likely to be admitted to the Intensive Care Unit (ICU) and had a higher risk of invasive infection, including development of bacteraemia. MLVA identified two separate VIM-2 MBL-producing clones, responsible for outbreaks in the ICU but also affecting 10 other departments. This is the first reported outbreak of VIM-2 MBL-producing P. aeruginosa in The Netherlands. Once introduced, VIM-2 MBL-producing P. aeruginosa cause significant infections and are easily spread within the hospital setting.

Population-based study of the epidemiology and the risk factors for Pseudomonas aeruginosa bloodstream infection.

BACKGROUND: Detailed population-based data on the epidemiology of Pseudomonas aeruginosa bloodstream infections are sparse. We sought to describe the incidence rate, risk factors, and outcomes associated with P. aeruginosa bacteremia in a large Canadian health region. PATIENTS AND METHODS: A retrospective population-based surveillance for P. aeruginosa bacteremia was conducted in the Calgary Health Region (CHR, population:approx. 1.2 million) during the period from 2000 to 2006. RESULTS: A total of 284 incident cases of P. aeruginosa bacteremia were identified in CHR residents, corresponding to an annual incidence rate of 3.6/100,000.Nosocomial acquisition accounted for 45% of cases,healthcare-associated community onset for 34% of cases,and community-acquired (CA) cases for 21%. Relative to the general population, risk factors for blood stream infection included male sex, increasing age, hemodialysis,solid organ transplant, diagnosis of cancer, heart disease, HIV infection, diabetes mellitus, and/or chronic obstructive airway disease (COPD). Overall mortality was 29%. Factors associated with mortality in univariate analysis included pulmonary focus of infection and co-morbidities, including chronic liver disease, substance abuse, heart disease, COPD, and cancer, and increased with the burden of co-morbidities. Despite those patients with CA disease having fewer co-morbidities,they had a significantly higher mortality rate than either healthcare-associated cases or nosocomial cases(RR 1.88, p = 0.05). CONCLUSIONS: This study documents that P. aeruginosa bacteremic disease is responsible for a significant burden of illness in general populations and identifies those groups at increased risk of infection and subsequent mortality. This information can be used to identify those individuals likely to benefit from empiric anti-pseudomonal therapies.

Detection of AmpC beta-lactamases in Escherichia coli, Klebsiella spp., Salmonella spp. and Proteus mirabilis in a regional clinical microbiology laboratory.

There are currently no standardized diagnostic tests available for the reliable detection of AmpC beta-lactamases in Klebsiella spp., Escherichia coli, Proteus mirabilis and Salmonella spp. A study was designed to evaluate a confirmation disk test using cefotetan (CTT) and cefoxitin (FOX) with phenylboronic acid (PBA). It also investigated the most suitable screening concentrations of FOX, ceftriaxone (CRO) and ceftazidime (CAZ) for the detection of AmpC beta-lactamases. A total of 126 control (consisting of 11 laboratory and 115 well-characterized clinical strains) and 29,840 non-repeat clinical isolates were included. FOX with PBA used in a confirmation test and CRO and CAZ as screening agents were found to be unreliable. FOX at >or= 32 mg/L was the best screening agent and CTT with PBA was the best confirmation test. Of the clinical isolates 635 (2%) were found to be resistant to cefoxitin (MIC >or= 32 ug/mL) and 332 (52%) were AmpC positive. E. coli was the most common organism with AmpC beta-lactamases and was mostly present in urines from community patients. It is recommended that laboratories use FOX at 32 mg/L as a screening agent and perform a disk test with CTT and PBA to confirm the presence of an AmpC cephalosporinase in isolates of Klebsiella spp., E. coli, Salmonella spp. and P. mirabilis. This approach is convenient, practical and easy to incorporate into the workflow of a clinical laboratory. False-positive AmpC detection may occur with KPC-producing bacteria when inhibitor-based methods are used.

Incidence, risk factors and outcomes of Escherichia coli bloodstream infections in a large Canadian region.

Although Escherichia coli is the most common cause of bloodstream infection, its epidemiology has not been well defined in non-selected populations. We sought to describe the incidence of risk factors for, and outcomes associated with, E. coli bacteraemia. Population-based surveillance for E. coli bacteraemia was conducted in the Calgary Health Region (population 1.2 million) during the period 2000-2006. In total, 2368 episodes of E. coli bacteraemia were identified for an overall annual population incidence of 30.3/100 000; 15% were nosocomial, 32% were healthcare-associated community-onset and 53% were community-acquired bacteraemias. The very young and the elderly were at highest risk for E. coli bacteraemia. Sixty per cent of the episodes occurred in females (relative risk 1.5; 95% CI 1.4-1.6). Dialysis, solid organ transplantation and neoplastic disease were the most important risk factors for acquiring E. coli bacteraemia. Rates of resistance to ampicillin, trimethoprim-sulphamethoxazole, gentamicin, ciprofloxacin, cefazolin and ceftriaxone increased significantly during the period 2000-2006. The case-fatality rate was 11% and the annual population mortality rate was 2.9/100 000. Increasing age, ciprofloxacin resistance, non-urinary focus and a number of comorbid illnesses were independently associated with an increased risk of death, and community acquisition and urinary focus were associated with a lower risk of death. This study documents the major burden of illness associated with E. coli bacteraemia and identifies groups at increased risk for acquiring and dying from these infections. The emergence of ciprofloxacin resistance and its adverse effect on patient outcome is a major concern.

Metallo-beta-lactamase-producing Pseudomonas aeruginosa isolated from a large tertiary centre in Kenya.

This study was designed to characterize the beta-lactamase content of carbapenem-resistant Pseudomonas aeruginosa isolates recovered during 2006 and 2007 in a large tertiary-care centre in Nairobi, Kenya. Molecular characterization was done using PCR and sequencing, and typing was performed using pulsed-field gel electrophoresis (PFGE). In total, 416 P. aeruginosa isolates were obtained during that period, of which 57 (13.7%) were resistant to carbapenems. All carbapenem-resistant isolates tested positive for metallo-beta-lactamase (MBL) production. All MBL isolates produced VIM-2 with two types of integron structures. PFGE identified three clonally related groups of VIM-2-producing P. aeruginosa, including a pan-resistant clone that was responsible for nosocomial outbreaks during 2006 and 2007 in the intensive-care unit. These findings suggest that continuous molecular surveillance needs to be performed to monitor the spread within the hospital of this pan-resistant strain. This study is the first report of VIM-2-producing P. aeruginosa from the African continent.

Population-based laboratory surveillance for Serratia species isolates in a large Canadian health region.

A population-based laboratory surveillance was conducted during a six-year period to define the incidence, demographic risk factors for acquisition, and anti-microbial susceptibilities of Serratia species isolates. A total of 715 incident Serratia species isolates were identified for an annual incidence of 10.8 per 100,000 residents; bacteremic disease occurred in 0.9 per 100,000 residents annually. The incidence increased with advancing age and males were at the highest risk. Ninety-two percent of the isolates were Serratia marcescens, and the majority (65%) of incident Serratia species isolates were of community onset. Ninety-five percent of isolates were susceptible to ciprofloxacin, 98% to gentamicin, 98% to trimethoprim/sulfamethoxazole, and >99% to imipenem. No yearly increase in resistance was observed. Serratia species isolation is most commonly of community onset and older patients and males are at increased risk. Despite reports of increasing resistance among Serratia species, the incidence in our region remains at a low stable rate.

Community-onset urinary tract infections: a population-based assessment.

BACKGROUND: Although multiple studies have investigated community-onset urinary tract infections (UTI), population-based data are lacking. We therefore conducted population-based laboratory surveillance in order to define the incidence, demographic risk factors, etiology, and antimicrobial susceptibilities of community onset UTI in a large Canadian region. METHODS: Laboratory surveillance for all community onset UTIs among residents of the Calgary Health Region (population approximately 1.2 million) was conducted during 2004/2005. Repeated positive samples within a 1-month period and those infections first cultured more than 2 days after admission to a hospital were excluded. RESULTS: A total of 40,618 episodes of community onset UTI occurred among 30,851 residents for an overall annual incidence of 17.5 per 1,000. Seventy-four percent of the cultures were submitted from ambulatory patients, 18% from hospitalized patients within the first 2 days of admission, and 9% from nursing home residents. Females were at significantly increased risk as compared to males (30.0 vs 5.0 per 1,000, RR 5.98; 95% CI, 5.81-6.15; p < 0.0001) as were the very young and very old. The most common infecting organisms were Escherichia coli (70%), Klebsiella pneumoniae (7%) and Enterococcus species (6%). Overall resistance rates among first isolates per patient tested were 14% for trimethoprim/sulfamethoxazole, 8% for cefazolin, 7% for nitrofurantoin, 6% for ciprofloxacin, 4% for gentamicin, and 2% for ceftriaxone although rates differed significantly based on sending location and patient age. CONCLUSION: This study provides novel information on the epidemiology of community-onset UTIs in a non-selected Canadian population. The occurrence, etiology, and resistance rates of community onset UTI differ significantly among definable population groups.

Population-based laboratory surveillance for tribe Proteeae isolates in a large Canadian health region.

The tribe Proteeae comprises the genera Proteus, Morganella and Providencia. Few studies have specifically investigated the epidemiology of infections caused by the Proteeae, and none has been conducted in a large non-selected population. The present study was a population-based laboratory surveillance in the Calgary Health Region (population 1.2 million), Canada during 2000-2005 that aimed to define the incidence, demographical risk-factors for acquisition and antimicrobial susceptibilities of Proteeae isolates. In total, 5047 patients were identified from whom Proteeae isolates were obtained (an annual incidence of 75.9/100 000), with females and the elderly being at highest risk. Incidence rates were 64.8, 7.7 and 3.4/100,000/year for the genera Proteus, Morganella and Providencia, respectively. Overall, 85% of infections were community-onset, and the overall rate of bacteraemic disease was 2.0/100,000. Compared with other species, Proteus mirabilis occurred at a much higher frequency, especially among females, and was less likely to be isolated from hospital-onset infections or to be part of a polymicrobial infection. Among isolates from community-onset infections, Providencia spp. were less likely to be from outpatients and more likely to be from nursing home residents. There were low overall rates of resistance to ciprofloxacin (4%) and gentamicin (5%), with Prot. mirabilis generally being the most susceptible. Members of the Proteeae were isolated frequently in both the community and hospital settings, but were infrequent causes of invasive disease. The occurrence, demographical risk-factors and microbiology of Proteeae isolates varied according to the individual species.

Treatment of infections caused by metallo-beta-lactamase-producing Pseudomonas aeruginosa in the Calgary Health Region.

This study reviewed 56 patients with significant metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa infections between May 2002 and March 2004 to identify features associated with mortality. Immunosuppression (p 0.002), bacteraemia (p 0.08) and inadequate antimicrobial therapy (p <0.001) were associated with death. Among those patients treated with adequate therapy, the use of multiple drug treatment regimens (two or three active agents) was associated with a non-significant two-fold increase in survival (p 0.45). Further prospective studies are warranted to determine the optimal treatment of MBL-producing P. aeruginosa infections.

Development and clinical validation of a molecular diagnostic assay to detect CTX-M-type beta-lactamases in Enterobacteriaceae.

Enterobacterial isolates producing CTX-M beta-lactamases have recently emerged worldwide in the community and hospital settings. Because of the significant public health implications, the spread of organisms producing CTX-M enzymes merits close monitoring with enhanced surveillance efforts. A molecular diagnostic assay using two different sets of primers simultaneously for the detection of all bla(CTX-M)-like beta-lactamase genes was developed. This assay repeatedly demonstrated 100% sensitivity, specificity and positive and negative predictive values for detecting different CTX-M enzymes in well-characterised strains that included producers of VEB-, TEM- and SHV-type extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC enzymes. The majority (132/240; 55%) of ESBL-producing enterobacterial isolates recovered in the Calgary Health Region during 2003 and 2004 were positive for bla(CTX-M) genes, including 81 (61%) positive for the CTX-M-9 group, 49 (37%) for the CTX-M-1 group, and two (2%) for the CTX-M-2 group. The CTX-M-specific PCR assay was reproducible and easy to use. It can be introduced in a clinical or reference laboratory to track and monitor the spread of organisms producing CTX-M enzymes in the community and hospital settings.