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1.
Antimicrob Agents Chemother ; 67(12): e0013623, 2023 12 14.
Article in English | MEDLINE | ID: mdl-37966229

ABSTRACT

Staphylococcus aureus is the most prevalent cystic fibrosis (CF) pathogen. Several phenotypes are associated with worsened CF clinical outcomes including methicillin-resistance and small-colony-variants. The inoculum effect (IE) is characterized by reduced ß-lactam susceptibility when assessed at high inoculum. The IE associates with worse outcomes in bacteremia and other high-density infections, and may therefore be relevant to CF. The prevalence of IE amongst a CF cohort (age ≥18 years), followed from 2013 to 2016, was investigated. Yearly methicillin-sensitive S. aureus (MSSA) isolates were screened at standard (5 × 105 CFU/mL) and high (5 × 107 CFU/mL) inoculum against narrow-spectrum anti-Staphylococcal ß-lactams and those with anti-pseudomonal activity common to CF. A ≥ 4-fold increase in minimum inhibitory concentration between standard and high inoculum defined IE. Isolates underwent blaZ sequencing and genotyping and were compared against published genomes. Fifty-six percent (99/177) of individuals had MSSA infection. MSSA was observed at ≥105 CFU/mL in 44.8% of entry sputum samples. The prevalence of the IE was 25.0%-cefazolin; 13.5%-cloxacillin; 0%-meropenem; 1.0%-cefepime; 5.2%-ceftazidime; and 34.4%-piperacillin-tazobactam amongst baseline MSSA isolates assessed. blaZ A associated with cefazolin IE (P = 0.0011), whereas blaZ C associated with piperacillin-tazobactam IE (P < 0.0001). Baseline demographics did not reveal specific risk factors for IE-associated infections, nor were long-term outcomes different. Herein, we observed the IE in CF-derived MSSA disproportionally for cefazolin and piperacillin-tazobactam and this phenotype strongly associated with underlying blaZ genotype. The confirmation of CF being a high density infection, and the identification of high prevalence of MSSA with IE in CF supports the need for prospective pulmonary exacerbation treatment studies to understand the impact of this phenotype.


Subject(s)
Cystic Fibrosis , Staphylococcal Infections , Adult , Humans , Adolescent , Methicillin/pharmacology , Methicillin/therapeutic use , Cefazolin/pharmacology , Staphylococcus aureus/genetics , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Prospective Studies , Cystic Fibrosis/drug therapy , Staphylococcal Infections/drug therapy , Staphylococcal Infections/epidemiology , Monobactams/pharmacology , Piperacillin, Tazobactam Drug Combination/therapeutic use , Ceftazidime/pharmacology , beta Lactam Antibiotics , Microbial Sensitivity Tests
3.
J Hosp Infect ; 132: 62-72, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36473552

ABSTRACT

BACKGROUND: Solid organ and haematopoietic stem cell transplant recipients are more vulnerable to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) than non-transplant recipients due to immunosuppression, and may pose a continued transmission risk, especially within hospital settings. Detailed case reports including symptoms, viral load and infectiousness, defined by the presence of replication-competent viruses in culture, provide an opportunity to examine the relationship between clinical course, burden and contagiousness, and provide guidance on release from isolation. OBJECTIVES: To investigate the relationship between serial SARS-CoV-2 reverse transcription polymerase chain reaction (RT-PCR) cycle threshold (Ct) value or cycle of quantification value, or other measures of viral burden and the likelihood and duration of the presence of infectious virus based on viral culture, including the influence of age, sex, underlying pathologies, degree of immunosuppression, and/or vaccination on this relationship, in transplant recipients. METHODS: LitCovid, medRxiv, Google Scholar and the World Health Organization COVID-19 database were searched from 1st November 2019 to 26th October 2022. Studies reporting relevant data (results from serial RT-PCR testing and viral culture data from the same respiratory samples) for transplant recipients with SARS-CoV-2 infection were included in this systematic review: Methodological quality was assessed using five criteria, and the data were synthesized narratively and graphically. RESULTS: Thirteen case reports and case series reporting on 41 transplant recipients (22 renal, five cardiac, one bone marrow, two liver, one bilateral lung and 10 blood stem cell) were included in this review. A relationship was observed between proxies of viral burden and likelihood of shedding replication-competent SARS-CoV-2. Three individuals shed replication-competent viruses for >100 days after symptom onset. Lack of standardization of testing and reporting platforms precludes establishing a definitive viral burden cut-off. However, the majority of transplant recipients stopped shedding replication-competent viruses when the Ct value was >30 despite differences across platforms. CONCLUSIONS: Viral burden is a reasonable proxy for infectivity when considered within the context of the clinical status of each patient. Standardized study design and reporting are essential to standardize guidance based on an increasing evidence base.


Subject(s)
COVID-19 , Organ Transplantation , Humans , COVID-19/diagnosis , SARS-CoV-2 , Viral Load , Hematopoietic Stem Cells
4.
J Hosp Infect ; 131: 1-11, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36195200

ABSTRACT

BACKGROUND: The arrival of the Delta variant of SARS-CoV-2 was associated with increased transmissibility and illness of greater severity. Reports of nosocomial outbreaks of Delta variant COVID-19 in acute care hospitals have been described but control measures varied widely. AIM: Epidemiological investigation of a linked two-ward COVID-19 Delta variant outbreak was conducted to elucidate its source, risk factors, and control measures. METHODS: Investigations included epidemiologic analysis, detailed case review serial SARS-CoV-2 reverse transcriptase-polymerase chain reaction (RT-PCR) testing of patients and healthcare workers (HCWs), viral culture, environmental swabbing, HCW-unaware personal protective equipment (PPE) audits, ventilation assessments, and the use of whole genome sequencing (WGS). FINDINGS: This linked two-ward outbreak resulted in 17 patient and 12 HCW cases, despite an 83% vaccination rate. In this setting, suboptimal adherence and compliance to PPE protocols, suboptimal hand hygiene, multi-bedded rooms, and a contaminated vital signs cart with potential fomite or spread via the hands of HCWs were identified as significant risk factors for nosocomial COVID-19 infection. Sudden onset of symptoms, within 72 h, was observed in 79% of all Ward 2 patients, and 93% of all cases (patients and HCWs) on Ward 2 occurred within one incubation period, consistent with a point-source outbreak. RT-PCR assays showed low cycle threshold (CT) values, indicating high viral load from environmental swabs including the vital signs cart. WGS results with ≤3 SNP differences between specimens were observed. CONCLUSION: Outbreaks on both wards settled rapidly, within 3 weeks, using a `back-to-basics' approach without extraordinary measures or changes to standard PPE requirements. Strict adherence to recommended PPE, hand hygiene, education, co-operation from HCWs, including testing and interviews, and additional measures such as limiting movement of patients and staff temporarily were all deemed to have contributed to prompt resolution of the outbreak.


Subject(s)
COVID-19 , Cross Infection , Humans , SARS-CoV-2/genetics , COVID-19/diagnosis , COVID-19/epidemiology , COVID-19/prevention & control , Personal Protective Equipment , Disease Outbreaks/prevention & control , Hospitals , Cross Infection/epidemiology , Cross Infection/prevention & control , Vital Signs , Health Personnel
5.
J Hosp Infect ; 130: 63-94, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36115620

ABSTRACT

BACKGROUND: The role of fomites in the transmission of SARS-CoV-2 is unclear. AIM: To assess whether SARS-CoV-2 can be transmitted through fomites, using evidence from viral culture studies. METHODS: Searches were conducted in the World Health Organization COVID-19 Database, PubMed, LitCovid, medRxiv, and Google Scholar to December 31st, 2021. Studies that investigated fomite transmission and performed viral culture to assess the cytopathic effect (CPE) of positive fomite samples and confirmation of SARS-CoV-2 as the cause of the CPE were included. The risk of bias using a checklist modified from the modified Quality Assessment of Diagnostic Accuracy Studies - 2 (QUADAS-2) criteria was assessed. FINDINGS: Twenty-three studies were included. The overall risk of bias was moderate. Five studies demonstrated replication-competent virus from fomite cultures and three used genome sequencing to match fomite samples with human clinical specimens. The mean cycle threshold (CT) of samples with positive viral culture was significantly lower compared with cultured samples that returned negative results (standardized mean difference: -1.45; 95% confidence interval (CI): -2.00 to -0.90; I2 = 0%; P < 0.00001). The likelihood of isolating replication-competent virus was significantly greater when CT was <30 (relative risk: 3.10; 95% CI: 1.32 to 7.31; I2 = 71%; P = 0.01). Infectious specimens were mostly detected within seven days of symptom onset. One study showed possible transmission of SARS-CoV-2 from fomites to humans. CONCLUSION: The evidence from published studies suggests that replication-competent SARS-CoV-2 is present on fomites. Replication-competent SARS-CoV-2 is significantly more likely when the PCR CT for clinical specimens and fomite samples is <30. Further studies should investigate the duration of infectiousness of SARS-CoV-2 and the frequency of transmission from fomites.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , SARS-CoV-2/genetics , Fomites , COVID-19/diagnosis
7.
J Hosp Infect ; 124: 56-66, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35307506

ABSTRACT

BACKGROUND: Multi-drug-resistant organisms (MDROs) (meticillin-resistant Staphylococcus aureus, vancomycin-resistant enterococcus, MDR Acinetobacter spp., extended-spectrum beta-lactamase-producing Gram-negative organisms), Clostridioides difficile, viral respiratory pathogens and healthcare-associated infections (HAIs) are major concerns in medical facilities. AIMS: To assess environmental service workers' (ESWs) training, workload, numbers of surfaces and pieces of medical equipment cleaned per shift, how long they worked on each surface, how they cleaned, supervision, MDRO and HAI status, and measurement of effectiveness in reducing MDROs and HAIs in patients and on surfaces. METHODS: CINAHL, Cochrane CENTRAL Register of Controlled Trials, EMBASE, Medline and Scopus were searched from inception to 28th June 2021 (no language/date limits). Searches combined terms from three themes: (a) hospitals and acute/long-term care facilities (e.g. assisted living, long-term care facilities, nursing homes); (b) disinfectants (e.g. antisepsis, bleach, cleaning, copper plating of surfaces and copper impregnation of textiles, disinfection, decontamination, hydrogen peroxide, quaternary ammonium disinfectants, and ultraviolet rays); and (c) randomized controlled trials (RCTs). FINDINGS AND CONCLUSIONS: Fourteen cluster RCTs provided data on a series of interventions. Ten trials focused on reducing patient infections from MDROs/HAIs, and four found significant reductions in patient infection rates with strategies including bleach, quaternary ammonium detergents, ultraviolet light and hydrogen peroxide vapour. Minimal information was provided on ESW numbers, MDRO/HAI status, workload, numbers of surfaces and objects cleaned per shift, and effects of training on improving effectiveness. The findings suggest that there is a need for detailed evaluations of the effects of training and disinfection activities for individuals and teams of ESWs.


Subject(s)
Ammonium Compounds , Cross Infection , Disinfectants , Copper , Cross Infection/prevention & control , Disinfectants/pharmacology , Disinfection , Hospitals , Humans , Hydrogen Peroxide , Long-Term Care , Nursing Homes
8.
J Hosp Infect ; 122: 9-26, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34998912

ABSTRACT

BACKGROUND: Multiply drug-resistant organisms (MDROs) in hospitals and long-term care facilities (LTCFs) of particular concern include meticillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococcus, multidrug-resistant Acinetobacter species, and extended-spectrum ß-lactamase-producing organisms. Respiratory viruses include influenza and SARS-CoV-2. AIM: To assess effectiveness of cleaning and disinfecting surfaces in hospitals and LTCFs. METHODS: CINAHL, Cochrane CENTRAL Register of Controlled Trials, Embase, Medline, and Scopus searched inception to June 28th, 2021, no language restrictions, for randomized controlled trials (RCTs), cleaning, disinfection, hospitals, LTCFs. Abstracts and titles were assessed and data abstracted independently by two authors. FINDINGS: Of 14 cluster (c)-RCTs in hospitals and LTCFs, interventions in ten were focused on reducing patient infections of four MDROs and/or healthcare-associated infections (HAIs). In four c-RCTs patient MDRO and/or HAI rates were significantly reduced with cleaning and disinfection strategies including bleach-, quaternary ammonium detergent-, ultraviolet irradiation-, hydrogen peroxide vapour- and copper-treated surfaces or fabrics. Of three c-RCTs focused on reducing MRSA rates, one had significant results and one on Clostridioides difficile had no significant results. Heterogeneity of populations, methods, outcomes and data reporting precluded meta-analysis. Overall risk of bias assessment was low but high for allocation concealment, and GRADE assessment was low risk. No study assessed biofilms. CONCLUSION: Ten c-RCTs focused on reducing multiple MDROs and/or HAIs and four had significant reductions. Three c-RCTs reported only patient MRSA colonization rates (one significant reductions), and one focused on C. difficile (no significant differences). Standardized primary and secondary outcomes are required for future c-RCTs including detailed biofilm cleaning/disinfection interventions.


Subject(s)
COVID-19 , Cross Infection , Methicillin-Resistant Staphylococcus aureus , Virus Diseases , COVID-19/prevention & control , Cross Infection/prevention & control , Drug Resistance, Multiple, Bacterial , Hospitals , Humans , Long-Term Care , SARS-CoV-2
9.
Article in English | MEDLINE | ID: mdl-31149332

ABSTRACT

Background: Given global issues with antimicrobial resistance and a need to optimize antimicrobial usage, antimicrobial stewardship (AS) programs are becoming a necessary component of hospitals and are increasingly mandated worldwide. It is important to evaluate these programs with respect to relevant clinical outcomes. Methods: An AS program with a prospective audit and feedback service (PAF) of antimicrobial usage was initiated May 11, 2015 at our tertiary care center, for patients admitted under the hospitalist service. We conducted a retrospective matched cohort study. Patients assessed during the first year of this PAF were considered to be the exposed cohort and were compared to unexposed controls matched on gender, age and infectious diagnosis selected from patients who had been admitted under the hospitalist service prior to initiation of the PAF. Descriptive analysis was completed and a multivariate conditional logistic regression was performed to analyze differences between the exposed and control groups in terms of a composite endpoint of 30 day mortality, 30 day post hospital discharge mortality and hospital re-admission. Results: A total of 348 patients were assessed and received PAF suggestions during the first year were compared to 827 matched control patients who did not receive PAF suggestions. Of 707 PAF suggestions made, the most common was to stop an antimicrobial (23%). A significantly lower (20.7% vs 28.8%, p = 0.008) composite endpoint was found in the group exposed to the PAF (OR 0.71 95%CI 0.52-0.97). This difference persisted when only patients with PAF suggestions that were completely or partially accepted were considered (18.6% vs 28.5%, p = 0.001) but was no longer significant when patients who had their ASP suggestions declined were analyzed (30.2% vs 26.7%, p = 0.610). Conclusions: In this retrospective cohort study, patient admissions in which PAF recommendations were accepted had better clinical outcomes than matched historical controls managed in the absence of this AS service.


Subject(s)
Antimicrobial Stewardship , Hospitalization/statistics & numerical data , Patient Admission/statistics & numerical data , Patient Readmission/statistics & numerical data , Adult , Aged , Anti-Bacterial Agents/therapeutic use , Female , Hospital Mortality , Hospitalists , Humans , Length of Stay , Logistic Models , Male , Middle Aged , Research Design , Retrospective Studies
10.
J Hosp Infect ; 103(1): 85-91, 2019 Sep.
Article in English | MEDLINE | ID: mdl-30991081

ABSTRACT

In a population-based, five-year retrospective cohort study of 5304 adult patients with hospital-acquired Clostridioides difficile infection across Alberta (N=101 hospitals), 30-day all-cause and attributable mortality were 12.2% and 4.5%, respectively. Patients >75 years of age had the highest odds of attributable mortality (odds ratio (OR) 9.34, 95% confidence interval (CI) 2.92-29.83) and largest difference in mean length of stay (11.7 days, 95% CI 8.2-15.2). A novel finding was that elevated white blood cell count at admission was associated with reduced attributable mortality (OR 0.67, 95% CI 0.50-0.90) which deserves further study. Advancing age was incrementally and significantly associated with all outcomes.


Subject(s)
Clinical Decision Rules , Clostridium Infections/diagnosis , Clostridium Infections/mortality , Cross Infection/diagnosis , Cross Infection/mortality , Length of Stay , Leukocyte Count , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Alberta/epidemiology , Clostridium Infections/epidemiology , Clostridium Infections/pathology , Cross Infection/epidemiology , Cross Infection/pathology , Female , Humans , Male , Middle Aged , Prognosis , Retrospective Studies , Survival Analysis , Young Adult
11.
Article in English | MEDLINE | ID: mdl-30181869

ABSTRACT

Background: Antimicrobial stewardship programs (ASPs) have been shown to reduce inappropriate antimicrobial use and its consequences. However, these programs lack legislative requirements in many places and it can be difficult to determine what human resources are required for these programs and how to create a business case to present to hospital administrators for program funding. The objectives of the current paper were to review legislative requirements and outline human resource requirements for ASPs, and to create a base business case for ASPs. Methods: A working group of antimicrobial stewardship experts from across Canada met to discuss the necessary components for creation of a business case for antimicrobial stewardship. A narrative review of the literature of the regulatory requirements and human resource recommendations for ASPs was conducted. Informed by the review and using a consensus decision-making process, the expert working group developed human resource recommendations based on a 1000 bed acute care health care facility in Canada. A spreadsheet based business case model for ASPs was also created. Results: Legislative and /or regulatory requirements for ASPs were found in 2 countries and one state jurisdiction. The literature review and consensus development process recommended the following minimum human resources complement: 1 physician, 3 pharmacists, 0.5 program administrative and coordination support, and 0.4 data analyst support as full time equivalents (FTEs) per 1000 acute care beds. Necessary components for the business case model, including the human resource requirements, were determined to create a spreadsheet based model. Conclusions: There is evidence to support the negative outcomes of inappropriate antimicrobial use as well as the benefits of ASPs. Legislative and /or regulatory requirements for ASPs are not common. The available evidence for human resource recommendations for ASPs using a narrative review process was examined and a base business case modelling scenario was created. As regulatory requirements for ASPs increase, it will be necessary to create accurate business cases for ASPs in order to obtain the necessary funding to render these programs successful.


Subject(s)
Antimicrobial Stewardship , Cross Infection/drug therapy , Cross Infection/microbiology , Emergency Medical Services , Anti-Infective Agents/pharmacology , Anti-Infective Agents/therapeutic use , Antimicrobial Stewardship/legislation & jurisprudence , Antimicrobial Stewardship/methods , Health Planning Guidelines , Humans , Models, Theoretical
12.
Epidemiol Infect ; 145(2): 254-265, 2017 01.
Article in English | MEDLINE | ID: mdl-27760576

ABSTRACT

An outbreak of Legionnaires' disease occurred in an inner city district in Calgary, Canada. This outbreak spanned a 3-week period in November-December 2012, and a total of eight cases were identified. Four of these cases were critically ill requiring intensive care admission but there was no associated mortality. All cases tested positive for Legionella pneumophila serogroup 1 (LP1) by urinary antigen testing. Five of the eight patients were culture positive for LP1 from respiratory specimens. These isolates were further identified as Knoxville monoclonal subtype and sequence subtype ST222. Whole-genome sequencing revealed that the isolates differed by no more than a single vertically acquired single nucleotide variant, supporting a single point-source outbreak. Hypothesis-based environmental investigation and sampling was conducted; however, a definitive source was not identified. Geomapping of case movements within the affected urban sector revealed a 1·0 km common area of potential exposure, which coincided with multiple active construction sites that used water spray to minimize transient dust. This community point-source Legionnaires' disease outbreak is unique due to its ST222 subtype and occurrence in a relatively dry and cold weather setting in Western Canada. This report suggests community outbreaks of Legionella should not be overlooked as a possibility during late autumn and winter months in the Northern Hemisphere.


Subject(s)
Disease Outbreaks , Genotype , Legionella pneumophila/classification , Legionella pneumophila/genetics , Legionnaires' Disease/epidemiology , Aged , Antigens, Bacterial/urine , Bacteriological Techniques , Canada/epidemiology , Female , Genomics , Humans , Legionella pneumophila/isolation & purification , Legionnaires' Disease/microbiology , Male , Middle Aged , Molecular Epidemiology , Molecular Typing , Sputum/microbiology , Surveys and Questionnaires , Urban Population
13.
Antimicrob Agents Chemother ; 60(3): 1794-800, 2016 Jan 11.
Article in English | MEDLINE | ID: mdl-26824951

ABSTRACT

This study describes 3 different blaNDM-1 genetic platforms in 3 different species obtained from the same patient who was directly transferred to an institution in Calgary, Alberta, Canada, following a prolonged hospital stay in India. The blaNDM-1 in the Escherichia coli isolate was located on a 176-kb IncA/C plasmid contained within an ISCR1 region. The blaNDM-1 in the Providencia rettgeri isolate was located on a 117-kb IncT plasmid contained within Tn3000, while the blaNDM-1 in the Pseudomonas aeruginosa isolate was located on the chromosome within an ISCR3 region. This report highlights the plasticity of the genetic regions and environments associated with blaNDM-1. To the best of our knowledge, this is the first report of P. aeruginosa with blaNDM-1 identified in North America and the first report of blaOXA-181 in P. rettgeri. The P. aeruginosa isolate belonged to the international high-risk sequence type 654 clone and was nonsusceptible to colistin. This case emphasizes the need for the use of appropriate infection prevention and control measures and vigilant screening for carbapenem-resistant Gram-negative bacteria in patients with a history of travel to areas of endemicity, such as the Indian subcontinent.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Colistin/therapeutic use , Drug Resistance, Bacterial/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Providencia/drug effects , Providencia/genetics , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics , beta-Lactamases/genetics , Aged , Canada , Carbapenems/therapeutic use , Escherichia coli/isolation & purification , Humans , India , Male , Microbial Sensitivity Tests , Plasmids/genetics , Providencia/isolation & purification , Pseudomonas aeruginosa/isolation & purification
14.
Antimicrob Agents Chemother ; 60(3): 1258-63, 2015 Dec 07.
Article in English | MEDLINE | ID: mdl-26643346

ABSTRACT

Enterobacteriaceae with blaNDM-7 are relatively uncommon and had previously been described in Europe, India, the United States, and Japan. This study describes the characteristics of Enterobacteriaceae (Klebsiella pneumoniae [n = 2], Escherichia coli [n = 2], Serratia marcescens [n = 1], and Enterobacter hormaechei [n = 1] isolates) with blaNDM-7 obtained from 4 patients from Calgary, Canada, from 2013 to 2014. The 46,161-bp IncX3 plasmids with blaNDM-7 are highly similar to other blaNDM-harboring IncX3 plasmids and, interestingly, showed identical structures within the different isolates. This finding may indicate horizontal transmission within our health region, or it may indicate contact with individuals from areas of endemicity within the hospital setting. Patients infected or colonized with bacteria containing blaNDM-7 IncX3 plasmids generate infection control challenges. Epidemiological and molecular studies are required to better understand the dynamics of transmission, the risk factors, and the reservoirs for bacteria harboring blaNDM-7. To the best of our knowledge, this is the first report of S. marcescens and E. hormaechei with blaNDM-7.


Subject(s)
Enterobacteriaceae/drug effects , Enterobacteriaceae/genetics , Plasmids/genetics , beta-Lactamases/genetics , Alberta/epidemiology , Bacterial Proteins/genetics , Enterobacteriaceae Infections/epidemiology , Enterobacteriaceae Infections/microbiology , Female , High-Throughput Nucleotide Sequencing/methods , Hospitals , Humans , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Male , Microbial Sensitivity Tests
15.
BMC Microbiol ; 15: 215, 2015 Oct 16.
Article in English | MEDLINE | ID: mdl-26474751

ABSTRACT

BACKGROUND: The application of molecular based diagnostics in sepsis has had limited success to date. Molecular community profiling methods have indicated that polymicrobial infections are more common than suggested by standard clinical culture. A molecular profiling approach was developed to investigate the propensity for polymicrobial infections in patients predicted to have bacterial sepsis. RESULTS: Disruption of blood cells with saponin and hypotonic shock enabled the recovery of microbial cells with no significant changes in microbial growth when compared to CFU/ml values immediately prior to the addition of saponin. DNA extraction included a cell-wall digestion step with both lysozyme and mutanolysin, which increased the recovery of terminal restriction fragments by 2.4 fold from diverse organisms. Efficiencies of recovery and limits of detection using Illumina sequencing of the 16S rRNA V3 region were determined for both viable cells and DNA using mock bacterial communities inoculated into whole blood. Bacteria from pre-defined communities could be recovered following lysis and removal of host cells with >97% recovery of total DNA present. Applying the molecular profiling methodology to three septic patients in the intensive care unit revealed microbial DNA from blood had consistent alignment with cultured organisms from the primary infection site providing evidence for a bloodstream infection in the absence of a clinical lab positive blood culture result in two of the three cases. In addition, the molecular profiling indicated greater diversity was present in the primary infection sample when compared to clinical diagnostic culture. CONCLUSIONS: A method for analyzing bacterial DNA from whole blood was developed in order to characterize the bacterial DNA profile of sepsis infections. Preliminary results indicated that sepsis infections were polymicrobial in nature with the bacterial DNA recovered suggesting a more complex etiology when compared to blood culture data.


Subject(s)
Bacteria/classification , Bacteria/genetics , Blood/microbiology , Coinfection/diagnosis , DNA, Bacterial/genetics , Molecular Diagnostic Techniques/methods , Sepsis/diagnosis , Adolescent , Adult , Aged , Aged, 80 and over , Cohort Studies , Coinfection/microbiology , Female , Humans , Male , Middle Aged , RNA, Ribosomal, 16S/genetics , Sepsis/microbiology , Young Adult
16.
East Mediterr Health J ; 19 Suppl 1: S39-47, 2013.
Article in English | MEDLINE | ID: mdl-23888794

ABSTRACT

Viruses account for the majority of the acute respiratory tract infections (ARIs) globally with a mortality exceeding 4 million deaths per year. The most commonly encountered viruses, in order of frequency, include influenza, respiratory syncytial virus, parainfluenza and adenovirus. Current evidence suggests that the major mode of transmission of ARls is through large droplets, but transmission through contact (including hand contamination with subsequent self-inoculation) and infectious respiratory aerosols of various sizes and at short range (coined as "opportunistic" airborne transmission) may also occur for some pathogens. Opportunistic airborne transmission may occur when conducting highrisk aerosol generating procedures and airborne precautions will be required in this setting. General infection control measures effective for all respiratory viral infections are reviewed and followed by discussion on some of the common viruses, including severe acute respiratory syndrome (SARS) coronavirus and the recently discovered novel coronavirus.


Subject(s)
Cross Infection/prevention & control , Delivery of Health Care/methods , Infection Control/methods , Respiratory Tract Infections/prevention & control , Acute Disease , Adenovirus Infections, Human/prevention & control , Adenovirus Infections, Human/transmission , Coronavirus Infections/prevention & control , Coronavirus Infections/transmission , Humans , Influenza, Human/prevention & control , Influenza, Human/transmission , Internationality , Paramyxoviridae Infections/prevention & control , Paramyxoviridae Infections/transmission , Respiratory Syncytial Virus Infections/prevention & control , Respiratory Syncytial Virus Infections/transmission , Respiratory Tract Infections/transmission , Severe Acute Respiratory Syndrome/prevention & control , Severe Acute Respiratory Syndrome/transmission
18.
J Antimicrob Chemother ; 68(7): 1505-9, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23524466

ABSTRACT

OBJECTIVES: Vancomycin-resistant enterococci (VRE) can be associated with serious bacteraemia. The focus of this study was to characterize the molecular epidemiology of VRE from bacteraemia cases that were isolated from 1999 to 2009 as part of Canadian Nosocomial Infection Surveillance Program (CNISP) surveillance activities. METHODS: From 1999 to 2009, enterococci were collected from across Canada in accordance with the CNISP VRE surveillance protocol. MICs were determined using broth microdilution. PCR was used to identify vanA, B, C, D, E, G and L genes. Genetic relatedness was examined using multilocus sequence typing (MLST). RESULTS: A total of 128 cases of bacteraemia were reported to CNISP from 1999 to 2009. In 2007, a significant increase in bacteraemia rates was observed in western and central Canada. Eighty-one of the 128 bacteraemia isolates were received for further characterization and were identified as Enterococcus faecium. The majority of isolates were from western Canada (60.5%), followed by central (37.0%) and eastern (2.5%) Canada. Susceptibilities were as follows: daptomycin, linezolid, tigecycline and chloramphenicol, 100%; quinupristin/dalfopristin, 96.3%; high-level gentamicin, 71.6%; tetracycline, 50.6%; high-level streptomycin, 44.4%; rifampicin, 21.0%; nitrofurantoin, 11.1%; clindamycin, 8.6%; ciprofloxacin, levofloxacin and moxifloxacin, 1.2%; and ampicillin, 0.0%. vanA contributed to vancomycin resistance in 90.1% of isolates and vanB in 9.9%. A total of 17 sequence types (STs) were observed. Beginning in 2006 there was a shift in ST from ST16, ST17, ST154 and ST80 to ST18, ST412, ST203 and ST584. CONCLUSIONS: The increase in bacteraemia observed since 2007 in western and central Canada appears to coincide with the shift of MLST STs. All VRE isolates remained susceptible to daptomycin, linezolid, chloramphenicol and tigecycline.


Subject(s)
Bacteremia/epidemiology , Cross Infection/epidemiology , Enterococcus faecium/classification , Gram-Positive Bacterial Infections/epidemiology , Vancomycin Resistance , Adolescent , Adult , Aged , Aged, 80 and over , Bacteremia/microbiology , Canada/epidemiology , Child , Child, Preschool , Cross Infection/microbiology , DNA, Bacterial/genetics , Enterococcus faecium/drug effects , Enterococcus faecium/genetics , Enterococcus faecium/isolation & purification , Female , Genes, Bacterial , Gram-Positive Bacterial Infections/microbiology , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Epidemiology , Multilocus Sequence Typing , Polymerase Chain Reaction , Young Adult
19.
Eur J Clin Microbiol Infect Dis ; 32(1): 33-42, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22898726

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) strains from different geographic areas have different genetic backgrounds, suggesting independent clonal evolutions. To better understand the virulence of MRSA strains and the relationship to their clonal and geographic origins, we undertook an analysis of epidemiologic, molecular, and virulence characteristics of a large number of MRSA isolates from geographically diverse origins, in a Caenorhabditis elegans infection model. A total of 99 MRSA isolates collected between 1993 and 2010 at the Geneva University Hospitals from diverse global origins were characterized with Panton-Valentine leukocidin (PVL), toxic shock syndrome toxin (TSST), accessory gene regulator (agr) group, staphylococcal cassette chromosome mec (SCCmec), S. aureus protein A (spa), multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE) typing. Epidemiologic data were provided from clinical records. The bacterial virulence was tested in a C. elegans host model. The inter-relationships of epidemiological/molecular characteristics in association with nematocidal activities were analyzed with univariate and two-factor analysis of variance (ANOVA). Community-associated MRSA (CA-MRSA) strains were more virulent than hospital-associated MRSA (HA-MRSA), with higher nematocidal activities in CA-MRSA strains (0.776 vs. 0.506, p = 0.0005). All molecular characteristics (PVL, TSST, spa, SCCmec, MLST, and PFGE types) showed a significant association with nematocidal activities on univariate analysis (p < 0.005). PVL was not a significant predictor after adjusting for genomic backgrounds using spa, MLST, or PFGE typing. The dominant CA-MRSA strains in North America showed higher nematocidal activities than strains from other regions (p < 0.0001). Strains with global origins containing distinct genetic backgrounds have different virulence in the C. elegans model. Nematocidal activities were most highly correlated with SCCmec, spa, MLST, and PFGE typing, suggesting that genomic background rather than a single exotoxin characteristic was the most discriminating predictor of virulence.


Subject(s)
Caenorhabditis elegans/microbiology , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Staphylococcal Infections/microbiology , Animals , Disease Models, Animal , Electrophoresis, Gel, Pulsed-Field , Global Health , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Molecular Typing , Multilocus Sequence Typing , Survival Analysis , Virulence , Virulence Factors/genetics
20.
(East. Mediterr. health j).
in English | WHO IRIS | ID: who-118445

ABSTRACT

Viruses account for the majority of the acute respiratory tract infections [ARIs] globally with a mortality exceeding 4 million deaths per year. The most commonly encountered viruses, in order of frequency, include influenza, respiratory syncytial virus, parainfluenza and adenovirus. Current evidence suggests that the major mode of transmission of ARIs is through large droplets, but transmission through contact [including hand contamination with subsequent selfinoculation] and infectious respiratory aerosols of various sizes and at short range [coined as [opportunistic] airborne transmission] may also occur for some pathogens. Opportunistic airborne transmission may occur when conducting highrisk aerosol generating procedures and airborne precautions will be required in this setting. General infection control measures effective for all respiratory viral infections are reviewed and followed by discussion on some of the common viruses, including severe acute respiratory syndrome [SARS] coronavirus and the recently discovered novel coronavirus


Subject(s)
Infection Control , Acute Disease , Delivery of Health Care , Tuberculosis , Health Personnel , Caregivers , World Health Organization , Respiratory Tract Infections
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