Concordance between epidemiological evaluation of probability of transmission and whole genome sequence relatedness among hospitalized patients acquiring Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae.

OBJECTIVES: We aimed to evaluate the concordance between epidemiologically determined transmission and genetic linkage of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-Kp). METHODS: We included consecutive KPC-Kp carriers between December 2016 and April 2017 in a hospital endemic for KPC-Kp. We assessed epidemiological relatedness between patients by prospective investigations by the infection control team. The probability of epidemiological relatedness was classified into four groups: no suspected transmission, low, moderate and high probability of transmission. Whole-genome sequencing of isolates was performed. Genetic linkage between KPC-Kp isolates was expressed by distance between isolates in single nucleotide polymorphisms (SNPs). We established an SNP cut-off defining a different strain based on the reconstructed phylogenetic tree. We compared the epidemiological and genetic linkage of all isolates from all patients. RESULTS: The study included 25 KPC-Kp carriers with 49 isolates. SNP variance was available for 1129 crossed patient-isolate pairs. Genomic linkage, based on a cut-off of 80 SNPs to define related isolates, was found in 115/708 (16.2%) of isolates with no transmission suspected epidemiologically, 27/319 (8.5%) of low, 11/26 (42.3%) of moderate and 64/76 (84.2%) of high epidemiological transmission risk determination (p < 0.001 for trend). Similar results and significant trends were shown on sensitivity analyses using a lower SNP cut-off (six SNPs) and patient-isolate unique pairs, analysing the first isolate from each patient. CONCLUSIONS: While significant concordance between epidemiological and genomic transmission patterns was found, epidemiological investigations of transmission are limited by the possibility of unidentified transmissions or over-estimation of associations. Genetic linkage analysis is an important aid to epidemiological transmission assessment.

Efficacy of a hospital policy of selective digestive decontamination for carbapenem-resistant Enterobacterales carriers: prospective before-after study.

A single-centre interrupted time series quasi-experimental study was undertaken to assess whether a hospital policy of selective digestive decontamination (SDD, gentamicin/amikacin with neomycin) administered to carbapenem-resistant Enterobacterales (CRE) carriers would reduce the duration of carriage and contain the spread of CRE. No significant difference in time to CRE eradication was observed between the observation (12 months, 120 patients) and intervention (12 months, 101 patients) periods. No change in the trend of new in-hospital CRE acquisitions or bacteraemia during the intervention was detected. As such, administration of SDD to CRE carriers was not effective for the eradication of carriage or controlling in-hospital CRE transmissions.

Combining VITEK® 2 with colistin agar dilution screening assist timely reporting of colistin susceptibility.

OBJECTIVES: The rise in carbapenem resistance among Gram-negative bacteria has renewed interest in colistin. Recently, the EUCAST-CLSI Polymyxin Breakpoints Working Group declared that broth microdilution (BMD) is the only valid method for colistin susceptibility testing. BMD is not easily incorporated into the routine work of clinical laboratories, and usually this test is incorporated serially, resulting in delayed susceptibility reporting. We tested a strategy of combining VITEK® 2 with a 2 µg/mL colistin agar dilution (VITEK® 2/AD) screening plate to improve performance and time to reporting of colistin susceptibility. METHODS: Colistin susceptibility for 364 clinical isolates was determined by VITEK® 2/AD and compared with the reference standard BMD according to the ISO 20776-1:2007 and CLSI guidelines. The EUCAST colistin susceptibility breakpoint of ≤2 µg/mL was used. Escherichia coli NCTC 13846 served as quality control strain. Agreement, very major error (VME) and major error rates were determined using ISO 20776-2:2007. RESULTS: The VME rate for VITEK® 2 alone was 30.6% (15/49, 95% CI 18.3-45.4%), and was reduced to 10.2% (5/49, 95% CI 3.4-22.2%) using the VITEK® 2/AD combined testing. The combined testing had categorical agreement with BMD of 97% (354/364, 95% CI 95.0-98.7%), and a major error (ME) rate of 1.6% (5/315, 95% CI 0.5-3.7%). Using the combined testing, even against challenging strains, 349 (95.8%, 95% CI 93.3-97.7%) colistin susceptibility results could be reported, and only 15 isolates required further analysis by BMD. DISCUSSION: Our method is simple to apply and allows rapid reporting of colistin susceptibility.

The use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid bacterial identification in patients with smear-positive bacterial meningitis.

OBJECTIVES: To assess the potential of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) in rapid identification of bacteria from smear-positive cerebrospinal fluid (CSF) in a cohort of patients with meningitis. METHODS: Single-centre observational study, including adults and children with community-acquired or postneurosurgical bacterial meningitis. Meningitis was defined using established criteria. Samples of CSF that had a positive CSF Gram stain were directly examined by MALDI-TOF-MS. Identification was considered accurate when identical to the CSF culture or PCR results (species and genus level). Laboratory workers performing the MALDI-TOF-MS and interpreting its results were blinded to the direct smear results, except for the fact that it was positive. MALDI-TOF-MS results were not conveyed to clinicians. RESULTS: MALDI-TOF-MS was tested on 44 CSF samples; ten samples were obtained from patients with community-acquired meningitis, and 34 samples were from patients with postneurosurgical meningitis. The assay identified bacteria correctly in 17/21 of the samples with Gram-negative rods observed on the direct smear, all obtained from patients who had undergone neurosurgery, (sensitivity 81%, 95% CI 64.2%-97.7%). In the postneurosurgical group, Gram-positive cocci were identified correctly in only 1/11 (9.1%) of the samples, and Candida species were not identified in two samples. Among patients with community-acquired meningitis, the assay did not identify Streptococcus pneumoniae in eight of eight samples, Neisseria meningitidis in one sample (1/1), and Streptococcus agalactiae in one sample (1/1). CONCLUSIONS: We found MALDI-TOF-MS to be useful in the rapid identification of Gram-negative rods directly from smear-positive CSF samples, but not of Gram-positive bacteria.

The pyogenic potential of the different Streptococcus anginosus group bacterial species: retrospective cohort study.

Streptococcus anginosus Group (SAG) bacteria are common causes of pyogenic infections (PIs). We examined the association between SAG species and the presence of a PI through a retrospective, observational, cohort study, between the years 2009 and 2015. All adults with clinically significant SAG infections in one hospital in Israel were assessed for association between SAG species and the presence of a PI defined as an abscess, empyema, or deep/organ space surgical site infection. Risk factors for PI were assessed using multivariate backward stepwise logistic regression analysis. We identified 263 patients with significant SAG infections, 182 (69%) of which were caused by S. anginosus, 45 (17·1%) by S treptococcus constellatus and 36 (13·7%) by S treptococcus intermedius. The mean age of the patients was 56·8 ± 19·1 years. PIs were identified among 160 (60%) of the patients and were mostly non-bacteraemic (147/160, 91·8%), while most non-PI patients had bacteraemia (70/103, 68%). S. anginosus and S. constellatus were associated with a significantly lower incidence of PI than S. intermedius, OR 0·18 (95% CI 0·06-0·53) and 0·14 (0·04-0·48), respectively. Patients with PI were younger and, in general, had less co-morbidities. S. intermedius was associated with pyogenic non-bacteraemic infections, while S. anginosus and S. constellatus were associated with bacteraemia with no abscess or empyema formation. These data may indicate differences in virulence mechanisms of these SAG bacteria.

The association between infection control interventions and carbapenem-resistant Enterobacteriaceae incidence in an endemic hospital.

BACKGROUND: Israel experienced a national outbreak of carbapenem-resistant Enterobacteriaceae (CRE) starting in 2006. AIM: To assess the association between infection control (IC) interventions implemented in a referral hospital in Israel and CRE incidence. METHODS: Retrospective quasi-experimental study of prospectively collected data. CRE incidence, defined as the number of patients newly acquiring CRE in surveillance or clinical samples per 100,000 hospital-days, was plotted quarterly between 2005 and 2016. IC interventions were applied at different time-points throughout this period. Data were collected on IC staffing, number of rectal surveillance cultures, and carbapenem consumption. Autocorrelated segmented linear regression analysis was used to assess the time-points at which a significant change in the CRE incidence trend occurred, and the association between the timing of IC intervention implementation and observed CRE trends was assessed. Trends between time-points were expressed as quarterly percent change (QPC) with 95% confidence intervals (CIs). FINDINGS: Between 2005 and 2008, CRE incidence increased significantly (QPC: 19.7%; CI: 11.5-28.4), reaching a peak of 186.6 new acquisitions per 100,000 hospital-days. From mid-2011 until the end of follow-up, there was a significantly decreasing incidence trend (QPC: -4.5; CI: -6.4 to -2.5). Cohorting of patients, screening of contacts and high-risk patients on admission were insufficient to control the epidemic. Improved hand hygiene compliance, cohorting with dedicated nursing staff, addition of regular screening in high-risk departments, and carbapenem restriction were required. Decreasing CRE incidence was observed with an infectious diseases/IC staffing of 1.2-1.5 per 100 beds and 20,000-36,000 yearly CRE surveillance samples. CONCLUSION: A multi-faceted hospital-wide intervention programme is required to control CRE in hospital settings.

Detection and characterization of carbapenemase-producing Enterobacteriaceae in wounded Syrian patients admitted to hospitals in northern Israel.

Since 2013, four hospitals in northern Israel have been providing care for Syrian nationals, primarily those wounded in the ongoing civil war. We analyzed carbapenemase-producing Enterobacteriaceae (CPE) isolates obtained from these patients. Isolate identification was performed using the VITEK 2 system. Polymerase chain reaction (PCR) was performed for the presence of bla KPC, bla NDM, and bla OXA-48. Susceptibility testing and genotyping were performed on selected isolates. During the study period, 595 Syrian patients were hospitalized, most of them young men. Thirty-two confirmed CPE isolates were grown from cultures taken from 30 patients. All but five isolates were identified as Klebsiella pneumoniae and Escherichia coli. Nineteen isolates produced NDM and 13 produced OXA-48. Among a further 29 isolates tested, multilocus sequence typing (MLST) showed that ST278 and ST38 were the major sequence types among the NDM-producing K. pneumoniae and OXA-48-producing E. coli isolates, respectively. Most were resistant to all three carbapenems in use in Israel and to gentamicin, but susceptible to colistin and fosfomycin. The source for bacterial acquisition could not be determined; however, some patients admitted to different medical centers were found to carry the same sequence type. CPE containing bla NDM and bla OXA-48 were prevalent among Syrian wounded hospitalized patients in northern Israel. The finding of the same sequence type among patients at different medical centers implies a common, prehospital source for these patients. These findings have implications for public health throughout the region.

A multicenter study of the clonal structure and resistance mechanism of KPC-producing Escherichia coli isolates in Israel.

Little is known about the molecular epidemiology of Klebsiella pneumoniae carbapenemase-producing Escherichia coli (KPCEC). We aimed to describe the clonal structure and resistance mechanisms of KPCEC in a multicenter study. The study included 88 isolates from four medical centres in Israel: Tel Aviv Medical Center (n = 17), Laniado Medical Center (n = 12), Sha'are-Zedek Medical Center (n = 38), and Rambam Medical Center (n = 21). Twelve (14%) KPCEC were from clinical sites and 86% from surveillance cultures. The clonal structure was studied by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and was highly diverse, with 79 and 45 different PFGE types and STs, respectively. The most common clones were ST-131 and ST-410, identified in 21 isolates (23%). Dominant clonal complexes (CCs) were CC131 (n = 16), CC410 (n = 14), CC10 (n = 17), and CC-69 (n = 6). The blaKPC-2 and blaKPC-3 genes were identified in 68 and 20 isolates, respectively. All isolates were non-susceptible to ertapenem; 16 (18%) and 35 (40%) isolates were susceptible (minimal inhibitory concentration ≤1 mg/L) to imipenem and meropenem, respectively. Isolates were susceptible to colistin, amikacin, ciprofloxacin, gentamicin, and trimethoprim-sulfamethoxazole in 100%, 87%, 28%, 27%, and 21% of the cases, respectively. blaKPC-Harbouring plasmids from Tel Aviv Medical Center as well as from six CC-131 isolates from the other centres were studied by Inc and pMLST typing. Sixteen of the 20 blaKPC2-harbouring plasmids were of identical type, IncN-pMLST ST-15. In conclusion, the clonal structure of KPCEC in Israel is characterized by the predominance of known international extended-spectrum ß-lactamase-producing clones and by high intra- and inter-institutional diversity. This suggests that in Israel, clonal spread does not play a major role in the dissemination of KPCEC.

Predominance of Gram-negative bacilli among patients with catheter-related bloodstream infections.

We evaluated changes in the epidemiology of catheter-related bloodstream infections (CRBSIs) between 1996 and 2012 in a tertiary care centre in Israel. The cohort included 1754 episodes of CRBSI. The incidence of CRBSIs decreased throughout the study period, whereas 30-day mortality following bacteraemia increased. There was a linear shift toward predominance of Gram-negative bacilli throughout the study period (p for trend<0.001). In 1996, 68% (68/100) of CRBSIs were caused by Gram-positive cocci, whereas in 2012 77.8% (28/26) were caused by Gram-negative bacilli. The shift towards Gram-negative CRBSIs and the associated mortality mandates that empirical treatment for CRBSIs be directed by local epidemiology.