Soft tissue infections in Belgian rugby players due to Streptococcus pyogenes emm type 81.
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Biomedical subjects
Publications and source records attributed to H Goossens.
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The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for Mycoplasma pneumoniae (n=9) and Chlamydia pneumoniae (n=5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine M. pneumoniae-positive throat swabs, the mean difference in Ct values between NucliSens easyMAG extraction and QIAGEN extraction was -2.26 (range, -5.77 to +0.60); for the detection of five C. pneumoniae-positive throat swabs, the average difference in Ct values between the two methods was -3.38 (range, -6.62 to -2.02); and for the detection of cytomegalovirus in 24 blood samples, the mean difference in Ct values between the two methods was -0.95 (range, -5.51 to +1.68). The NucliSens easyMAG is considerably easier to perform, efficiently extracts nucleic acids from throat swabs and whole blood, is automated, and has high throughput.
Isothermal nucleic acid sequence-based amplification (NASBA) was applied to detect Legionella 16S rRNA. The assay was originally developed as a Legionella pneumophila conventional NASBA assay with electrochemiluminescence (ECL) detection and was subsequently adapted to a L. pneumophila real-time NASBA format and a Legionella spp. real-time NASBA using molecular beacons. L. pneumophila RNA prepared from a plasmid construct was used to assess the analytical sensitivity of the assay. The sensitivity of the NASBA assay was 10 molecules of in vitro wild type L. pneumophila RNA and 0.1-1 colony-forming units (CFU) of L. pneumophila. In spiked respiratory specimens, the sensitivity of the NASBA assays was 1-10000 CFU of L. pneumophila serotype 1 depending on the background. After dilution of the nucleic acid extract prior to amplification, 1-10 CFU of L. pneumophila serotype 1 could be detected with both detection methods. Finally, 27 respiratory specimens, well characterized by culture and PCR, collected during a L. pneumophila outbreak, were tested by conventional and real-time NASBAs. All 11 PCR positive samples were positive by conventional NASBA, 9/11 and 10/11 were positive by L. pneumophila real-time NASBA and Legionella spp. real-time NASBA, respectively.
OBJECTIVES: To collect reliable, comparable and publicly available data on hospital use of antibiotics in Europe aggregated at the national level (1997-2002). METHODS: Consumption data of systemic antibiotics in Anatomical Therapeutic Chemical (ATC) class J01 were collected and expressed in defined daily doses (DDD) per 1000 inhabitants per day. Valid data for 2002 were available for 15 countries, and 6 year trends for 10 countries. Comparison with ambulatory care (AC) consumption data was possible in 14 countries. RESULTS: In 2002, median national hospital antibiotic consumption in Europe was 2.1 DDD/1000 inhabitants/day in Europe, ranging from 3.9 in Finland and France to 1.3 in Norway and Sweden. Hospital care (HC) consumption as a proportion of total antibiotic consumption ranged from 17.8% to 6.4%. The consumption of hospital-specific antibiotics ranged from 0.43 DDD/1000 inhabitants/day in Greece and 0.08 in Sweden. Six-year trends in consumption were stable, except for rising co-amoxiclav exposure and more rapid market penetration of new antibiotics (e.g. levofloxacin) in some countries. There was a strong, positive correlation between the extent of antibiotic use in AC and in HC (Spearman coefficient 0.745; P = 0.002), both for overall use and for use of five main classes (not macrolides and 'others'). In contrast to AC consumption no substantial seasonal variation in consumption was observed. CONCLUSIONS: It was cumbersome but feasible to collect ecological data on hospital antibiotic consumption in a set of 15 European countries on a retrospective basis, illustrating substantial cross-national variations in the extent and distribution of exposure to antibiotics in hospital care.
According to the results of the European Surveillance of Antimicrobial Consumption (ESAC), outpatient antibiotic use in Croatia in 2001 (17.6 DDD per 1000 inhabitants per day) was comparable to the median use of 24 European countries, which were able to deliver valid data.
This study evaluated retrospectively the efficacy of treatment with cefepime vs. a carbapenem, in combination with amikacin or ciprofloxacin, for seriously-ill patients infected with ESBL-producing Enterobacter aerogenes who were admitted to an intensive care unit. Forty-four episodes of infection were investigated in 43 patients: 21 treated with cefepime; 23 with a carbapenem. The two treatment groups did not differ statistically in terms of age, APACHE II scores, and infection sites, but the average duration of antibiotic exposure was significantly shorter in the cefepime group (8.5 days vs. 11.4 days; p 0.04). Clinical improvement was seen in 62% of patients receiving cefepime vs. 70% of patients receiving a carbapenem (p 0.59). Bacteriological eradication was achieved in 14% of patients receiving cefepime vs. 22% of patients receiving a carbapenem (p 0.76). The 30-day mortality rates related to infection were 33% in the cefepime group and 26% in the carbapenem group (p 0.44). Thus, outcome parameters did not differ significantly between the two groups. Nevertheless, a statistically significant increase in failure to eradicate ESBL-producing E. aerogenes was observed as the MICs of cefepime rose (p 0.017). Pulsed-field gel electrophoresis revealed three distinct clones, but one predominant clone harbouring the bla(TEM-24) gene was associated with most (42/44) of the episodes of infection. It was concluded that cefepime may be an alternative agent for therapy of severe infections caused by TEM-24 ESBL-producing E. aerogenes, although further studies are required to confirm these observations.
This observational study describes the antimicrobial susceptibility testing (AST) methods and interpretive criteria used in European hospitals during 2001, focusing specifically on detection of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Of 263 hospitals that took part in the ARPAC study, 192 submitted data on AST. Of these, 89% (n = 170) routinely used a disk-diffusion AST method, 43% (n = 82) used a semi-automated method, and 70% (n = 135) routinely determined MICs. Hospitals in southern Europe were less likely to use disk-diffusion, but were more likely to use a semi-automated method (p <0.001). In total, 173 (90%) interpreted AST results using CLSI breakpoints; 30% of these detected MRSA using unmodified CLSI disk-diffusion methods, while 35% used the unmodified CLSI agar-screening method for MRSA; 41% and 30% adhered to unmodified CLSI methodology for disk-diffusion and agar-screening, respectively, to detect VRE. Some of the modifications made may have greatly reduced the ability of the tests to detect MRSA/VRE. For example, 20% of respondents used excessively high incubation temperatures and 13% used inadequate incubation times to detect MRSA by disk-diffusion, and 28% used Mueller-Hinton agar instead of brain-heart infusion agar in VRE screening plates. The majority of respondents stated that they followed CLSI guidelines, but a high proportion had modified the CLSI methods for detecting MRSA and VRE, which may compromise clinical management and antimicrobial resistance surveillance.
A set of 118 strains of the species Lactobacillus rhamnosus was collected, including probiotic strains, research strains with potential probiotic properties, food starter cultures, and human isolates. The majority of the strains were collected from companies, hospitals, or culture collections or were obtained after contacting authors who reported clinical case studies in the literature. The present work aimed to reveal the genotypic relationships between strains of these diverse sources. All strains were initially investigated using fluorescent amplified fragment length polymorphism (FAFLP) with three different primer combinations. Numerical analysis of FAFLP data allowed (i) confirmation of the identification of all strains as members of L. rhamnosus and (ii) delineation of seven stable intraspecific FAFLP clusters. Most of these clusters contained both (potentially) probiotic strains and isolates of human origin. For each of the clusters, strains of different sources were selected for pulsed-field gel electrophoresis (PFGE) of macrorestriction fragments obtained with the enzymes NotI and AscI. Analysis of PFGE data indicated that (i) some (potentially) probiotic strains were indistinguishable from other probiotic strains, suggesting that several companies may use duplicate cultures of the same probiotic strain, and (ii) in a number of cases human isolates from sterile body sites were indistinguishable from a particular probiotic strain, suggesting that some of these isolates may be reisolations of commercial strains.
Five hundred seventeen consecutive nasopharyngeal aspirates were collected between October 1998 and May 1999 for episodes of acute respiratory tract infections in children presenting at the University Hospital of Antwerp. Culture and nucleic acid amplification techniques--nucleic acid sequence-based amplification (NASBA) and reverse transcription-PCR (RT-PCR)--were applied to detect rhinoviruses (RVs). Other respiratory viruses were detected by immunofluorescence (IF) analysis of the specimens and IF analysis of shell vial cultures. Among the 517 specimens, 219 viral agents were identified. They were, in decreasing order, rhinoviruses (93 [18.0%]), respiratory syncytial virus (76 [14.7%]), adenoviruses (16 [3.1%]), influenza viruses (15 [2.9%]), enteroviruses (15 [2.9%]), and herpes simplex virus (4 [0.8%]). For the evaluation of rhinovirus detection, culture positivity and/or a positive reaction in the two independent amplification methods was used as an expanded "gold standard." Based on this standard, the sensitivity, specificity, positive predictive value, and negative predictive value of culture were 44.7, 100, 100, and 99.8%, and those of NASBA and RT-PCR were 85.1, 98.3, 83.3, and 98.5% and 82.9, 93.4, 55.7, and 98.2%, respectively. NASBA and RT-PCR produced comparable results and were significantly more sensitive than virus culture. RVs showed the highest incidence in acute respiratory tract infections in children.
The quality performance of laboratories for the detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae by two quality control (QC) exercises with a 2-year interval was investigated. For the 2002 QC exercise, specimens were spiked with M. pneumoniae at concentrations of 5,000, 500, 50, and 0 color-changing units (CCU)/100 microl. The limit of detectability was 50 CCU/100 microl. Therefore, this concentration was omitted from the 2004 panel and was excluded from the analysis. In 2002, 2 out of 12 participants obtained 100% correct results, 2 out of 12 produced false-positive results, and 10 out of 12 had between 0 out of 9 and 8 out of 9 correct positive results. In 2004, correct results were obtained in 15 out of 18 tests, and no false-positive results were reported. In 2002, specimens were spiked with C. pneumoniae at concentrations of 490, 49, 4.9, and 0 inclusion-forming units/100 microl (IFU/100 microl). In the 2004 panel, samples spiked with a lower dilution of 0.49 IFU/100 microl were added to the panel. For the C. pneumoniae QC, correct results were produced in 12 out of 16 and 13 out of 18 tests in 2002 and in 2004, respectively. Both multiplex PCR and nucleic acid sequence-based amplification (NASBA) formats scored a smaller number of samples positive than the monoplex reactions.
Isothermal nucleic acid sequence-based amplification (NASBA) was applied to the detection of Chlamydophila pneumoniae 16S rRNA by using the NucliSens basic kit (bioMérieux, Boxtel, The Netherlands). The assay was originally developed as a conventional NASBA assay with electrochemiluminescence detection and was subsequently adapted to a real-time NASBA format by using a molecular beacon. C. pneumoniae RNA prepared from a plasmid construct was used to assess the analytical sensitivity of the assay. The sensitivity of the NASBA assay was 10 molecules of in vitro wild-type C. pneumoniae RNA and 0.1 inclusion-forming unit (IFU) of C. pneumoniae. In spiked respiratory specimens, the sensitivity of the C. pneumoniae NASBA assay varied between 0.1 and 1 IFU/100 mul sample, depending on the type of specimen. Finally, conventional and real-time NASBA were applied to respiratory specimens previously tested by PCR. A 100% concordance between the test results was obtained.
To obtain better insights into the possible exchange of resistance genes between human and animal streptococci, the sequences of the erm (B) genes of streptococcal isolates from humans, pigs, pork carcasses, chickens, and calves were compared. Identical erm (B) gene sequences were present in strains from humans, pigs, pork carcasses, and calves. During in vitro mating experiments, the erm (B) gene was exchanged between porcine Streptococcus suis and human S. pneumoniae, S. pyogenes, and S. oralis strains. The presence of different tetracycline resistance genes and the int Tn 1545 gene was determined in animal streptococci carrying the erm (B) gene. Although tet(M) and int Tn 1545 genes were detected in 24% of the porcine and pork carcass streptococcal strains, the tet(O) gene was the predominant tetracycline resistance gene in these strains (81%). The latter gene was co-transferred with the erm (B) gene from porcine S. suis strains to human streptococci in the mating experiments. These results show that, identical erm (B) gene sequences were present in animal and human streptococci and that transfer of the erm (B) gene from porcine S. suis to human streptococci and vice versa is possible, but probably occurs at a low frequency.
There is a need to improve surveillance systems in order to recognise emerging threats, both in the community and in hospitals, in a timely manner. The laboratory arm of surveillance must be complemented by hospital and primary care components. We also need more training at all levels: undergraduate and postgraduate medical training, specialist training, as well as continuing professional development schemes. Core training programmes for infectious disease specialists and medical microbiologists must be established. The contribution of general practitioners, infection specialists, microbiology laboratories, and public health specialists/agencies to response systems should be defined as clearly as possible and should be either empirically based or derived from scenario-guided calamity models. The success of surveillance hinges on sufficient long-term resources and dedicated coordination. The European Centre for Disease Prevention and Control (ECDC) may assume this role. However, structural and administrative constraints of the diverse healthcare systems throughout Europe may represent an impediment to a harmonised response. Also, the funding of the ECDC cannot cover the operational demands. The ECDC may well work as a small coordinating unit; however, without a laboratory-based scientific status, it remains doubtful whether the ECDC will achieve the competence and authority needed for effective leadership in healthcare. The ESCMID, together with other national and international scientific societies, should put emphasis on the fact that infectious diseases have not received the necessary degree of attention from governments in the European region. The ESCMID should strengthen its role in harmonising and supporting the highest standards of training in the infection disciplines.
BACKGROUND: Europe is a continent with strong public healthcare systems, but diverging antibiotic policies and resistance patterns. AIMS: To describe the performance and methodological approach in a retrospective data collection effort (1997-2001), through an international network of surveillance systems, aiming to collect publicly available, comparable and reliable data on antibiotic use in Europe. METHODS: A central multidisciplinary management team co-ordinated a network of national representatives, liasing with national data providers and bodies responsible for antibiotic policy. The data collected were screened for bias, using a checklist. We focused on detection bias in sample and census data; errors in assigning medicinal product packages to the Anatomical Therapeutic Chemical Classification (ATC); errors in calculations of defined daily doses (DDD) per package; bias by over-the-counter sales and parallel trade; and bias in ambulatory care (AC)/hospital care (HC) mix. Datasets were corrected after national feedback, and classified as valid; valid but with minor bias; not valid. RESULTS: Of the 31 participating countries, 21 countries delivered AC data suitable for cross-national comparison (14 for all 5 years). Of these, 17 countries provided data on a quarterly basis for at least 1 year. For HC, 14 countries were able to deliver valid data (nine for all 5 years). A valid estimate of the total exposure of national populations to human antibiotic consumption could be made in 17 countries. CONCLUSION: In cross-national comparisons of antibiotic consumption in Europe, methodological rigour in correcting for various sources of bias and checking the validity of ATC/DDD assignment is needed.
Bacteria of the genus Acinetobacter are ubiquitous in nature. These organisms were invariably susceptible to many antibiotics in the 1970s. Since that time, acinetobacters have emerged as multiresistant opportunistic nosocomial pathogens. The taxonomy of the genus Acinetobacter underwent extensive revision in the mid-1980s, and at least 32 named and unnamed species have now been described. Of these, Acinetobacter baumannii and the closely related unnamed genomic species 3 and 13 sensu Tjernberg and Ursing (13TU) are the most relevant clinically. Multiresistant strains of these species causing bacteraemia, pneumonia, meningitis, urinary tract infections and surgical wound infections have been isolated from hospitalised patients worldwide. This review provides an overview of the antimicrobial susceptibilities of Acinetobacter spp. in Europe, as well as the main mechanisms of antimicrobial resistance, and summarises the remaining treatment options for multiresistant Acinetobacter infections.
An omnibus survey of microbiologists (n = 400) and a survey of participants (n = 49) in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) programme were conducted to determine the awareness and prevalence of extended-spectrum beta-lactamases (ESBLs), and the regularity and method of screening. Of the omnibus survey participants, 69% screened regularly for ESBLs, compared with 83% of MYSTIC participants. In both surveys, ESBLs were more common in Klebsiella pneumoniae (73% and 79%, respectively) and Escherichia coli (63% and 81%, respectively) than in other bacteria. The surveys demonstrated that awareness of, and testing for, ESBLs is inconsistent.
This paper describes an outbreak of Legionnaire's disease at Kapellen in Belgium among visitors of the annual fair. The investigation started on 13th November 1999 after a respiratory physician notified the health authorities of the province of Antwerp of presumptive cases of legionellosis. The annual commercial fair at Kapellen, a small town in northern Belgium, was held 10 days previously and attracted 50,000 visitors. Stand employees (professionals or volunteers), technical staff of the hall and visitors at the fair were affected cases. An exploratory case-control study was conducted to trace the source of the epidemic. To complete the inventory study and to evaluate other risk factors, a cohort study of exhibitors and staff was conducted. Ninety-three people met the case definition, 41 of whom were considered as confirmed, 14 as presumptive cases and 38 as possible/clinical cases. Five people died. Further testing at the reference laboratory confirmed all strains to be Legionella pneumophila serogroup 1. The sensitivity for culture was low (29.2%), and sensitivity for seroconversion was high (90.9%). For urinary antigen test, a sensitivity with Biotest EIA of 65.6% was found, and the sensitivity of polymerase chain reaction (PCR) was 85.7%. In all cases, the individual had visited the fair. Those individuals working in the central areas of the tent, near the aerosol-producing devices, were at higher risk of disease. Legionella was detected by PCR on swabs of the surfaces of the whirlpool. Although not fully proven, an aerosol-producing device was the most probable source of the outbreak.
A commercially available nucleic acid sequence-based amplification (NASBA) NucliSens Basic Kit (NBK) assay for the detection of Mycoplasma pneumoniae 16S rRNA in respiratory specimens was developed and compared to standard NASBA and PCR assays previously developed in our laboratory. The specificity and sensitivity of the NBK assay was comparable to the specificity and sensitivity of the corresponding standard NASBA assay. The NBK offers standardized reagents for the development of a NASBA assay for the detection of M. pneumoniae in respiratory specimens and is easily adaptable to other amplification targets.