Phenotypic and molecular characterizations of carbapenem-resistant Acinetobacter baumannii isolates collected within the EURECA study.

Multi-drug-resistant Acinetobacter baumannii isolates are key pathogens that contribute to the global burden of antimicrobial resistance. This study aimed to investigate the phenotypic and molecular characteristics of carbapenem-resistant A. baumannii (CRAB) isolates from the EURECA clinical trial. In total, 228 CRAB clinical strains were recovered from 29 sites in 10 European countries participating in the EURECA study between May 2016 and November 2018. All strains were reconfirmed centrally for identification and antimicrobial susceptibility testing, and were then subjected to DNA isolation and whole-genome sequencing (WGS), with analysis performed using BacPipe v.1.2.6. K and O typing was performed using KAPTIVE. Overall, 226 (99.1%) strains were confirmed as CRAB isolates. The minimum inhibitory concentration (MIC90) results of imipenem and meropenem were >16 mg/L. WGS showed that the isolates mainly harboured blaOXA-23 (n=153, 67.7%) or blaOXA-72 (n=70, 30.1%). Four blaOXA-72 isolates from Serbia co-harboured blaNDM-1. An IS5 transposase family element, ISAba31, was found upstream of the blaOXA-72 gene harboured on a small (~10-kb) pSE41030-EUR plasmid. The majority of isolates (n=178, 79.1%) belonged to international clone II. Strains belonging to the same sequence type but isolated in different countries or within the same country could be delineated in different clusters by core-genome multi-locus sequence typing (MLST). Whole-genome/core-genome MLST showed high diversity among the isolates, and the most common sequence type was ST2 (n=153, 67.7%). The EURECA A. baumannii strain collection represents a unique, diverse repository of carbapenem-resistant isolates that adds to the existing knowledge of A. baumannii epidemiology and resistance genes harboured by these strains.

Nosocomial outbreak of extended-spectrum ß-lactamase-producing Enterobacter cloacae among cardiothoracic surgical patients: causes and consequences.

BACKGROUND: Enterobacteriaceae are recognized as leading pathogens of healthcare-associated infections. AIM: To report the investigation of a nosocomial outbreak of extended-spectrum ß-lactamase-producing Enterobacter cloacae affecting cardiothoracic surgery patients in a Belgian academic hospital. METHODS: Cases were defined based on epidemiological and microbiological investigations, including molecular typing using repetitive element-based polymerase chain reaction and multi-locus sequence typing. Case-control studies followed by field evaluations allowed the identification of a possible reservoir, and the retrospective assessment of human and financial consequences. FINDINGS: Over a three-month period, 42 patients were infected or colonized by CTX-M-15-producing E. cloacae strains that belonged to the same clonal lineage. Acquisition mainly occurred in the intensive care unit (N = 23) and in the cardiothoracic surgery ward (N = 16). All but one patient had, prior to acquisition, undergone a cardiothoracic surgical procedure, monitored by the same transoesophageal echocardiography (TOE) probe in the operating room. Despite negative microbiological culture results, the exclusion of the suspected probe resulted in rapid termination of the outbreak. Overall, the outbreak was associated with a high mortality rate among infected patients (40%) as well as significant costs (€266,550). CONCLUSION: The outbreak was indirectly shown to be associated with the contamination of a manually disinfected TOE probe used per-operatively during cardiothoracic surgery procedures, because withdrawal of the putative device led to rapid termination of the outbreak.

Prospective evaluation of a high multiplexing real-time polymerase chain reaction array for the rapid identification and characterization of bacteria causative of nosocomial pneumonia from clinical specimens: a proof-of-concept study.

The purpose of this study was evaluation of the VAPChip assay based on the "Rapid-Array-PCR-technology" which targets 13 respiratory pathogens and 24 ß-lactam resistance genes directly on respiratory clinical specimens. The first step included analysis of 45 respiratory specimens in order to calibrate and determine the threshold for target genes. The second prospective step involved 85 respiratory samples from patients suspected of nosocomial pneumonia collected in two academic hospitals over an 8-month period. Results of the VAPChip assay were compared to routine methods. The first step showed a large proportion of positive signals for H. influenzae and/or S. pneumoniae. For identification, discrepancies were observed in seven samples. Thresholds were adapted and two probes were re-designed to create a new version of the cartridge. In the second phase, sensitivity and specificity of the VAPchip for bacterial identification were 72.9% and 99.1%, respectively. Seventy (82%) pathogens were correctly identified by both methods. Nine pathogens detected by the VAPChip were culture negative and 26 pathogens identified by culture were VAPChip negative. For resistance mechanisms, 11 probes were positive without identification of pathogens with an antimicrobial-susceptibility testing compatible by culture. However, the patient's recent microbiological history was able to explain most of these positive signals. The VAPChip assay simultaneously detects different pathogens and resistance mechanisms directly from clinical samples. This system seems very promising but the extraction process needs to be automated for routine implementation. This kind of rapid point-of-care automated platform permitting a syndromic approach will be the future challenge in the management of infectious diseases.

Increasing incidence of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in Belgian hospitals.

Carbapenemase-producing Enterobacteriaceae are increasingly reported worldwide. The aim of the study was to determine the incidence and molecular epidemiology of carbapenemase-producing (CP) Escherichia coli and Klebsiella pneumoniae (CP-E/K) in Belgium. Eleven hospital-based laboratories collected carbapenem non-susceptible (CNS) isolates of E. coli and K. pneumoniae detected in clinical specimens from January 2013 to December 2014. All CNS strains were tested for carbapenemase production and typed by multilocus sequence typing (MLST) for a 6-month period as part of the European Survey on Carbapenemase-Producing Enterobacteriaceae in Europe (EuSCAPE) structured survey. In addition, an equal number of carbapenem-susceptible isolates collected were preserved as a control group for risk factor analysis. The overall incidence rate of CP-E/K isolates in hospitals increased from 0.124 in 2013 to 0.223 per 1000 admissions in 2014. From November 2013 to April 2014, 30 CP K. pneumoniae [OXA-48 (n = 16), KPC (n = 13), OXA-427 (n = 1)] and five CP E. coli [OXA-48 (n = 3), NDM (n = 1), OXA-427 (n = 1)] isolates were detected in ten hospitals. The 16 OXA-48-producing K. pneumoniae strains were distributed into eight sequence types (STs), while the 13 KPC-producing K. pneumoniae clustered into three STs dominated by ST512 (n = 7) and ST101 (n = 5). Compared to controls, we observed among CP-E/K carriers significantly higher proportion of males, respiratory origins, previous hospitalization, nosocomial setting, and a significantly lower proportion of bloodstream infections. Our study confirms the rapid spread of CP-E/K in Belgian hospitals and the urgent need for a well-structured and coordinated national surveillance plan in order to limit their dissemination.

About three cases of ulceroglandular tularemia, is this the re-emergence of Francisella tularensis in Belgium?

Tularemia is a zoonosis caused by Francisella tularensis that can be transmitted by several ways to human being and cause different clinical manifestations. We report three clinical cases of tularemia with ulceroglandular presentation in young males acquired during outdoor activities in Southern Belgium. Confirmation of the diagnosis was established by serology. Only three cases of tularemia have been reported in Belgium between 1950 and 2012 by the National Reference Laboratory CODA-CERVA (Ref Lab CODA-CERVA) but re-emergence of tularemia is established in several European countries and F. tularensis is also well known to be present in animal reservoirs and vectors in Belgium. The diagnosis of tularemia has to be considered in case of suggestive clinical presentation associated with epidemiological risk factors.

Infection due to travel-related carbapenemase-producing Enterobacteriaceae, a largely underestimated phenomenon in Belgium.

BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) are emerging worldwide, representing a major threat for public health. Early CPE detection is crucial in order to prevent infections and the development of reservoirs/outbreaks in hospitals. In 2008, most of the CPE strains reported in Belgium were imported from patients repatriated from abroad. Actually, this is no longer the case. OBJECTIVES AND METHODS: A surveillance was set up in Belgian hospitals (2012) in order to explore the epidemiology and determinants of CPE, including the link with international travel/hospitalization. The present article describes travel-related CPE reported in Belgium. Different other potential sources for importation of CPE are discussed. RESULTS: Only 12% of all CPE cases reported in Belgium (2012-2013) were travel related (with/without hospitalization). This is undoubtedly an underestimation (missing travel data: 36%), considering the increasing tourism, the immigration from endemic countries, the growing number of foreign patients using scheduled medical care in Belgium, and the medical repatriations from foreign hospitals. The free movement of persons and services (European Union) contributes to an increase in foreign healthcare workers (HCW) in Belgian hospitals. Residents from nursing homes located at the country borders can be another potential source of dissemination of CPE between countries. Moreover, the high population density in Belgium can increase the risk for CPE-dissemination. Urban areas in Belgium may cumulate these potential risk factors for import/dissemination of CPE. CONCLUSIONS: Ideally, travel history data should be obtained from hospital hygiene teams, not from the microbiological laboratory. Patients who received medical care abroad (whatever the country) should be screened for CPE at admission.

Evaluation of several phenotypic methods for the detection of carbapenemase-producing Pseudomonas aeruginosa.

The purpose of this investigation was to compare several phenotypic methods, including combined disk tests (CDT) containing metallo-ß-lactamase (MBL) inhibitors or cloxacillin, and the Carba NP test for the detection of carbapenemase-producing Pseudomonas aeruginosa (CPPA). A new CDT using imipenem (10 µg) ± cloxacillin 4,000 µg and the Carba NP test were evaluated to detect CPPA. In addition, four commercially available combined disks containing a carbapenem and ethylene-diamine-tetra-acetic acid (EDTA) or dipicolinic acid (DPA) as the inhibitor were tested in order to detect MBL-positive P. aeruginosa. All these phenotypic methods were evaluated on 188 imipenem non-susceptible P. aeruginosa (CPPA, n = 75) isolates divided into 26 well-characterized collection strains and 162 non-duplicate clinical isolates referred to the national reference laboratory in 2013. For the total of 188 isolates tested, CDT containing EDTA or DPA displayed high sensitivities (99%) and specificities (95%) for detecting MBL-producing isolates. CDT with cloxacillin showed a sensitivity and specificity of 97%/96% compared to 88%/99% for the Carba NP test in order to detect CPPA. For the 162 clinical isolates, CDT containing EDTA or DPA displayed a high negative predictive value (NPV) (99%) for detecting MBL-producing isolates. CDT with cloxacillin showed an NPV of 98%, compared to 95% for the Carba NP test in order to detect CPPA. In our setting, CDT associating imipenem ± EDTA or ± DPA performed best for the detection of MBL-producing P. aeruginosa. Imipenem/imipenem-cloxacillin test yielded good NPV to exclude the presence of MBL in imipenem non-susceptible isolates.

Performance of different culture methods and of a commercial molecular assay for the detection of carbapenemase-producing Enterobacteriaceae in nursing homes and rehabilitation centers.

Over the last several years, carbapenemase-producing Enterobacteriaceae (CPE) have been increasingly detected not only among patients in acute care hospitals, but also in long-term care facilities. In this point prevalence survey, residents from three nursing homes and patients in one rehabilitation center were screened for asymptomatic intestinal carriage of CPE by rectal swabs. The first objective was to evaluate the hypothesis of the establishment of a CPE reservoir in a geriatric/chronic care population. Secondly, we evaluated the comparative performances of different culture methods (chromID(®) CARBA, chromID(®) OXA-48, MacConkey with temocillin/meropenem, ertapenem enrichment broth) and a commercial molecular assay (Check-Direct CPE). From the 257 included residents, only one had evidence for CPE carriage. From the rectal swabs of this resident, an OXA-48-producing Klebsiella pneumoniae could be isolated and was confirmed by a molecular assay both on the strain and on the rectal swab. The specificity of the different culture methods and Check-Direct CPE was at least 97 %. Neither enrichment broth nor prolonged incubation up to 48 h increased the yield of CPE. This point prevalence survey shows a low CPE prevalence of 0.39 %. Larger scaled studies are needed in order to confirm the role of chronic care settings as secondary CPE reservoirs and to adjust the infection control and prevention recommendations.

Prevalence and distribution of beta-lactamase coding genes in third-generation cephalosporin-resistant Enterobacteriaceae from bloodstream infections in Cambodia.

Resistance to third-generation cephalosporins in Gram-negative bacteria is emerging in Asia. We report the prevalence and distribution of extended-spectrum beta-lactamase (ESBL), AmpC beta-lactamase and carbapenemase-coding genes in cefotaxime-resistant Enterobacteriaceae isolates from bloodstream infections (BSI) in Cambodia. All Enterobacteriaceae isolated from BSI in adult patients at Sihanouk Hospital Centre of HOPE, Phnom Penh, Cambodia (2007-2010) were assessed. Antimicrobial susceptibility testing was carried out by disc diffusion and MicroScan according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Screening for ESBL, plasmidic AmpC and carbapenemase-coding genes was performed by multiplex polymerase chain reaction (PCR) sequencing assays. Identification of the ST131 clone was performed in all CTX-M-positive Escherichia coli, using PCR targeting the papB gene. Out of 183 Enterobacteriaceae, 91 (49.7 %) isolates (84 BSI episodes) were cefotaxime-resistant: E. coli (n = 68), Klebsiella pneumoniae (n = 17) and Enterobacter spp. (n = 6). Most episodes were community-acquired (66/84; 78.3 %). ESBLs were present in 89/91 (97.8 %) cefotaxime-resistant isolates: 86 (96.6 %) were CTX-M, mainly CTX-M-15 (n = 41) and CTX-M-14 (n = 21). CTX-M of group 1 were frequently associated with TEM and/or OXA-1/30 coding genes and with phenotypic combined resistance to ciprofloxacin, sulphamethoxazole-trimethoprim and gentamicin (39/50, 78.0 %). Plasmidic AmpC (CMY-2 and DHA-1 types) were found alone (n = 2) or in combination with ESBL (n = 4). Eighteen E. coli isolates were identified as B2-ST131-O25B: 11 (61.1 %) carried CTX-M-14. No carbapenemase-coding genes were detected. ESBL among Enterobacteriaceae from BSI in Cambodia is common, mainly associated with CTX-M-15 and CTX-M-14. These findings warrant urgent action for the containment of antibiotic resistance in Cambodia.

Reducing time to identification of positive blood cultures with MALDI-TOF MS analysis after a 5-h subculture.

Speeding up the turn-around time of positive blood culture identifications is essential in order to optimize the treatment of septic patients. Several sample preparation techniques have been developed allowing direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification of positive blood cultures. Yet, the hands-on time restrains their routine workflow. In this study, we evaluated an approach whereby MALDI-TOF MS identification without any additional steps was carried out on short subcultured colonies from positive blood bottles with the objective of allowing results reporting on the day of positivity detection. Over a 7-month period in 2012, positive blood cultures detected by 9 am with an automated system were inoculated onto a Columbia blood agar and processed after a 5-h incubation on a MALDI-TOF MicroFlex platform (Bruker Daltonik GmbH). Single-spotted colonies were covered with 1 µl formic acid and 1 µl matrix solution. The results were compared to the validated identification techniques. A total of 925 positive blood culture bottles (representing 470 bacteremic episodes) were included. Concordant identification was obtained in 727 (81.1 %) of the 896 monomicrobial blood cultures, with failure being mostly observed with anaerobes and yeasts. In 17 episodes of polymicrobic bacteremia, the identification of one of the two isolates was achieved in 24/29 (82.7 %) positive cultures. Routine implementation of MALDI-TOF MS identification on young positive blood subcultures provides correct results to the clinician in more than 80 % of the bacteremic episodes and allows access to identification results on the day of blood culture positivity detection, potentially accelerating the implementation of targeted clinical treatments.

Emergence and dissemination of multi-resistant Gram negative Enterobacteriaceae: lessons to be learnt from local and national surveillance programs in Belgium.

Infections caused by multi drug resistant bacteria (MDRB) constitutes an international health care problem. Since the year 2000, a longitudinal surveillance programme (LSP) and two multicentric surveys (100 hospitals, 826 isolates) were performed to monitor the emergence of MDRB in Belgium. The implementation of a LSP detected the emergence and spread of new types of ESBLs (CTX-M), mostly among community associated E. coli in the setting of a university hospital several years before the large spread and recognition in Belgium of a pathogenic E. coli CTX-M-15 (B2-O25:H4-ST131) pandemic clone (found in extra-intestinal virulent strains). This finding supports the progressive increase in Belgium of systemic infections including UTI caused by MDRB with limited therapeutical options. The real burden of the problem remains however, difficult to estimate in the absence of any surveillance network in Belgium to monitor the epidemiology of antimicrobial resistance in the community. The current Belgian national recommendations for the detection, surveillance, prevention and control of epidemics by ESBL-producing organisms and possibly other MDRBs (eg: Carbapenemase producing Enterobacteriaceae [CPE]) must be updated taking into accounts these new elements. A global coordinated network for antimicrobial surveillance resistance gathering experts (e.g: public health epidemiologists, representative of the national reference centres of antimicrobial resistance, field experts in infection control, infectious disease specialists, other clinicians and general practitioners) must be urgently implemented, including the longitudinal analysis of resistance in different ecosystems (human, animal, water and food).

Enteroaggregative Shiga toxin-producing Escherichia coli of serotype O104:H4 in Belgium and Luxembourg.

In 2011, a large outbreak of infections caused by Shiga toxin-producing Escherichia coli (STEC) O104:H4 occurred in Germany. This exceptionally virulent strain combined virulence factors of enteroaggregative E. coli (EAggEC) and STEC. After the outbreak only a few sporadic cases of infection with this rare serotype were reported, most of which were related to travel to the Middle East or North Africa. Here we describe two cases of enteroaggregative STEC (Agg-STEC) O104:H4 infection that occurred in Belgium in 2012 and 2013 respectively. In both cases travel in a Mediterranean country preceded the infection. The first strain was isolated from the stool of a 42-year-old woman presenting bloody diarrhoea, who had travelled to Tunisia the week before. The second case involves a 14-year-old girl who, upon her return from Turkey to Belgium, suffered from an episode of bloody diarrhoea and haemolytic uraemic syndrome. Extended typing of the isolates with pulsed field gel electrophoresis revealed that the strains were closely related, though not exactly the same as the 2011 outbreak strain. This report supports the previously made hypothesis that Agg-STEC has a human reservoir and might be imported by travellers coming from an area where the pathogen is endemic. Furthermore, it emphasizes the concern that these bacteria may cause future outbreaks as evenly virulent O104:H4 isolates seem to be widespread.

Epidemiological investigation of a nosocomial outbreak of multidrug-resistant Corynebacterium striatum at one Belgian university hospital.

During an 8-month period, 24 Corynebacterium striatum isolates recovered from lower respiratory tract specimens of 10 hospitalized patients were characterized. The organisms were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and by 16S rRNA gene sequencing. The cluster of C. striatum exclusively affected patients who had been admitted to an intensive care unit and/or subsequently transferred to one medium-size respiratory care unit. Prolonged duration of hospitalization, advanced stage of chronic obstructive pulmonary disease, recent administration of antibiotics and exposure to an invasive diagnostic procedure were the most commonly found risk factors in these patients. Seven patients were colonized and three infected. All strains displayed a similar broad spectrum resistance to antimicrobial agents, remaining susceptible to vancomycin only. Typing analysis by MALDI-TOF MS and by semi-automated repetitive sequence-based PCR (DiversiLab typing) showed that all outbreak-associated C. striatum isolates clustered together in one single type while they differed markedly from epidemiologically unrelated C. striatum isolates. Pulsed-field gel electrophoresis (PFGE) profiles revealed three distinct PFGE types among the C. striatum isolates associated with the outbreak while all external strains except one belonged to a distinct type. We conclude that C. striatum is an opportunistic nosocomial pathogen in long-term hospitalized patients and can be at the origin of major outbreaks. The routine use of MALDI-TOF MS greatly facilitated the recognition/identification of this organism in clinical samples and this technique could also offer the potential to be used as an easy and rapid epidemiological typing tool for outbreak investigation.

Carbapenemase-producing Enterobacteriaceae in Europe: a survey among national experts from 39 countries, February 2013.

The spread of carbapenemase-producing Enterobacteriaceae (CPE) is a threat to healthcare delivery, although its extent differs substantially from country to country. In February 2013, national experts from 39 European countries were invited to self-assess the current epidemiological situation of CPE in their country. Information about national management of CPE was also reported. The results highlight the urgent need for a coordinated European effort on early diagnosis, active surveillance, and guidance on infection control measures.

Probe ligation and real-time detection of KPC, OXA-48, VIM, IMP, and NDM carbapenemase genes.

The Check-MDR Carba test (Check-Points, Wageningen, Netherlands), which is based on specific molecular recognition of blaNDM, blaKPC, blaOXA-48, blaVIM, and blaIMP genes by DNA probe ligation and real-time PCR detection, was evaluated on 183 well-characterized Gram-negative rods. Representatives of the 5 gene families were accurately identified (specificities and sensitivities of 100%) within 4.5 hours. This test may be helpful to differentiate carbapenem resistance mediated by carbapenemases from those involving other mechanisms.

Validation of carbapenemase and extended-spectrum ß-lactamase multiplex endpoint PCR assays according to ISO 15189.

OBJECTIVES: To validate and accredit a set of three multiplex endpoint PCR assays, targeting the most important carbapenemase and minor extended-spectrum ß-lactamase (ESBL) resistance genes, according to the international ISO 15189 particular requirements for the quality and competence of medical laboratories. METHODS: Specific primers targeting ESBLs and carbapenemases were collected from the literature or designed internally. The multiplex PCRs were validated for sensitivity, specificity, intra- and inter-run reproducibility and accuracy by means of external quality control (EQC) using a collection of 137 characterized and referenced isolates. For each multiplex PCR assay, the presence of an extraction control ruled out false-negative results due to PCR inhibition or extraction faults. Amplicons were separated by capillary electrophoresis (QIAxcel system, Qiagen). The protocols and validation files were reviewed in the setting of an external audit conducted by the Belgian organization for accreditation (BELAC). RESULTS: Sensitivity, specificity and reproducibility for each targeted gene were 100%. All isolates from the three EQC panels were correctly identified by each PCR assay (accuracy 100%). The validation files were controlled by BELAC, and the PCR protocols were accepted as accredited according to ISO 15189. CONCLUSIONS: Three home-made multiplex PCRs targeting the major carbapenemases and four minor class A ESBL genes encountered in Gram-negative bacteria were accredited according to the ISO 15189 standards. This validation scheme could provide a useful model for laboratories aiming to accredit their own protocols.

Analytical validation of a novel high multiplexing real-time PCR array for the identification of key pathogens causative of bacterial ventilator-associated pneumonia and their associated resistance genes.

OBJECTIVES: Rapid diagnosis and appropriate empirical antimicrobial therapy before the availability of conventional microbiological results is of pivotal importance for the clinical outcome of ventilator-associated pneumonia (VAP). We evaluated the VAPChip, a novel, closed cartridge molecular tool aiming to identify directly from clinical samples and within a working day the principal bacteria causative of VAP as well as clinically relevant ß-lactam resistance genes. METHODS: The Real-time Array PCR for Infectious Diseases (RAP-ID) is a novel technology that combines multiplex PCR with real-time microarray detection. The VAPChip is a closed cartridge kit adapted to the RAP-ID instrument that targets 13 key respiratory pathogens causative of VAP and 24 relevant antimicrobial resistance genes that mediate resistance to ß-lactam agents, including extended-spectrum cephalosporins and carbapenems. Analytical validation of the VAPChip was carried out blindly on a collection of 292 genotypically characterized bacterial reference and clinical isolates, including 225 isolates selected on the basis of their species identification and antimicrobial resistance profiles and 67 bacterial isolates belonging to the oropharyngeal flora not targeted by the array. RESULTS: The limit of detection of the assay lies between 10 and 100 genome copies/PCR and the dynamic range is five orders of magnitude permitting at least semi-quantitative reporting of the results. Sensitivity, specificity and negative and positive predictive values ranged from 95.8% to 100% for species identification and detection of resistance genes. CONCLUSIONS: VAPChip is a novel diagnostic tool able to identify resistant bacterial isolates by RAP-ID technology. The results of this analytical validation have to be confirmed on clinical specimens.

External validation of the bacterial meningitis score in children hospitalized with meningitis.

UNLABELLED: The Bacterial Meningitis Score (BMS) is considered as the rule with the highest sensitivity to safely distinguish between aseptic and bacterial meningitis (BM). OBJECTIVE: The objective of our study was to evaluate the performance of the score and its usefulness for the clinician. METHOD: Retrospective analysis of two Belgian academic hospitals-based cohort studies. All consecutive children aged 29 days to 18 years admitted for acute meningitis between January 1996 and December 2008 was eligible. The BMS (risk of bacterial meningitis if seizure, positive cerebrospinal fluid (CSF) Gram staining, CSF protein level (3) 80 mg/dl, CSF neutrophil count 1,000/ mm3 or blood neutrophil count > or = 10,000/mm3) was applied to all patients with meningitis defined by CSF pleocytosis > 8 WBC/mm3. RESULTS: 174 patients were included in the final analysis of whom 26 (15%) had BM. Of the 93 patients categorized as having with no risk for BM (BMS score = 0), 2 patients had BM, one of which had petechial rash (negative predictive value 97.8%). BMS had a sensitivity of 92.3%. Risk of BM was significantly related to the BMS score: 6/147 (4%) patients with BMS < or = 1 had BM compared to 20/27 (74%) patients with BMS > 1. CONCLUSIONS: Our study reports a lower sensitivity of the BMS than observed in previous validation studies. We suggest to include the BMS in a decision tree aiming to optimize the ordering of laboratory investigations including viral and bacterial PCR testing in any child with CSF pleocytosis.

Clinical, virological and epidemiological assessment of 2009 influenza A (H1N1) pandemic in a Belgian university hospital.

BACKGROUND: Recommendations were applied before and during the Belgian pandemic (2009) H1N1 influenza wave at a university hospital (420 beds), for optimizing isolation processes and therapeutic management of possible and confirmed infected cases. METHODS: All patients presenting to the Emergency Department (ED) between August 1st and December 31st 2009 were screened for ILI symptoms, and were isolated for clinical assessment in case of positive screening. Patients categorized as possible influenza cases and who required hospitalization were isolated in dedicated wards. Specific diagnostic algorithms were implemented. Medical charts were retrospectively reviewed and matched with results of the microbiology laboratory. Patient's characteristics were analyzed, the contribution of laboratory diagnosis on therapy and lengh of stay (LOS) in isolation was also assessed. RESULTS: 310 patients out of 6068 had a positive screening for ILI, of these, 265 were retained as possible influenza cases and 139 required hospitalization. Twenty-eight children (8 requiring hospitalization) and 20 hospitalized adult patients had confirmed influenza infection. Five adult patients were admitted to the intensive care unit (ICU), 3 requiring extracorporeal membrane oxygenation (ECMO). There was no death related to the new influenza strain. The majority of confirmed patients were diagnosed during the Belgian epidemic wave, with a sensitivity of antigen detection of 50% in children and 35% in adults comparatively to real-time PCR (RT-PCR). CONCLUSIONS: The impact of (2009) H1N1 pandemic influenza remained limited, except for ICU patients requiring ECMO. Implementation of screening, isolation, and virological diagnosis processes led to significant improvement of patient management.