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H Goossens

Publications and source records attributed to H Goossens.

At least 91 records · Page 5Linked to original sources
PubMed1998

Molecular epidemiology of recent belgian isolates of Neisseria meningitidis serogroup B.

In Belgium an increase in the incidence of meningococcal disease has been noted since the early 1990s. Four hundred twenty clinical strains isolated during the period from 1990 to 1995, along with a set of 30 European reference strains, and 20 Dutch isolates were examined by random-primer and repetitive-motif-based PCR. A subset was investigated by multilocus enzyme electrophoresis and pulsed-field gel electrophoresis. The data were compared with results obtained by serotyping (M. Van Looveren, F. Carion, P. Vandamme, and H. Goossens, Clin. Microbiol. Infect. 4:224-228, 1998). Both phenotypic and molecular epidemiological data suggest that the lineage III of Neisseria meningitidis, first encountered in The Netherlands in about 1980, has been introduced in Belgium. The epidemic clone, as defined by oligonucleotide D8635-primed PCR, encompasses mainly phenotypes B:4:P1.4 and B:nontypeable:P1.4, but strains with several other phenotypes were also encountered. Therefore, serotyping alone would underestimate the prevalence of the epidemic clone.

Belgium↗
PubMed1998

An outbreak of Burkholderia cepacia with septicemia on a cardiology ward.

During a 3-day period, eight patients developed septicemia with Burkholderia cepacia. Heparin injection was found to be a risk factor. Heparin was diluted with dextrose solution, which was aspirated from a 1-L bag. B cepacia, genotypically identical to the blood isolates, was isolated from this bag.

Bacteremia↗
PubMed1997

Use of specialised isolation media for recognition and identification of Candida dubliniensis isolates from HIV-infected patients.

During a study of oral rinses of 130 HIV-infected individuals, both typical and atypical Candida albicans colonies were isolated from ten patients on a yeast differential medium. Typical Candida albicans colonies were light green; atypical colonies were dark green. Both types of colonies were germ tube-positive and produced chlamydospores. However, DNA fingerprinting of the atypical isolates with the Ca3 Candida albicans-specific probe showed that they belonged to the recently described species Candida dubliniensis. Candida dubliniensis colonies could also be differentiated from Candida albicans colonies on isolation plates by the absence of fluorescence of colonies on methyl blue-Sabouraud agar under Wood's light. Among other phenotypic characteristics, only the absence of intracellular beta-glucosidase activity reliably distinguished Candida albicans from Candida dubliniensis. Candida dubliniensis may be underreported in clinical samples because most currently used isolation and identification methods fail to recognize this yeast.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Diphtheria immunity in Flanders.

A serological survey to determine the immunity to diphtheria in the Flemish population was conducted according to the recommendations of the World Health Organization. Immunity to diphtheria was determined on a randomised, stratified sample (1679 serum samples) from an existing serum bank (4058 serum samples) representative of the Flemish population. All age groups between 0 and 100 years were included. A tissue (Vero cell) culture toxin neutralisation assay was used to measure serum diph-theria antitoxin concentrations. The results showed that 43% of the Flemish population was protected against diphtheria (antitoxin titre, > or = 0.1 IU/ml), while 32% was susceptible (antitoxin titre, < 0.01 IU/ml); for 25%, protection was of limited duration (antitoxin titre, > or = 0.01 IU/ml and < 0.1 IU/ml). The proportion of susceptible subjects showed a significant age-related increase, with the highest values in the 35 to 44 and 45 to 54 age groups (57.9% and 55.5%, respectively). These results emphasise the need for booster immunization of adults.

Adolescent↗
PubMed1997

Possible implication of sterile connecting device in contamination of pooled platelet concentrates.

Considering the possibility that a pooled random donor platelet concentrate could become contaminated by welding with a sterile connecting device, we undertook a study to determine the influence of pooling on the contamination rate. As a control group, apheresis platelets were examined. Bacteriological testing was done with a sensitive CO2 detecting culture system, the BacT/ Alert. Out of 1105 pooled platelet concentrates prepared by the buffy coat method, 15 (1.4%) were confirmed as contaminated, all with Staphylococcus epidermidis and two with a second bacterial species, i.e. Staphylococcus capitis and Propionibacterium acnes, respectively. Median detection time by the BacT/Alert was 23 h. Twelve pools of five units were contaminated, which is significantly more than the three contaminated pools of four units. On the other hand, the reuse of the welding wafers proved not be a risk factor for contamination. One welded tubing segment of a contaminated platelet concentrate failed the air leakage test, an incident which was 73 times more frequent than with the sterile platelet concentrates. We found five pooled platelet concentrates containing Staphylococci from which no bacteria could be grown from the individual buffy coats that had been pooled. We suggest the contamination here to have occurred after separation of the buffy coat from the whole blood, possibly during the welding process. Finally, none out of 378 apheresis platelet concentrates was contaminated. All our observations highlight the potential risk for contamination when making pooled platelet concentrates with a sterile connecting device. For this type of transfusion product, we advocate bacteriological screening of all units before release. The incubation time for the sterility test should, however, be limited to 36 h, if logistical problems with the availability of platelets are to be avoided.

Automation↗
PubMed1997

Evaluation of arbitrarily primed PCR analysis and pulsed-field gel electrophoresis of large genomic DNA fragments for identification of enterococci important in human medicine.

The increasing problems encountered with enterococcal nosocomial infections and the intrinsic and acquired resistance of the enterococci to different antimicrobial compounds highlight the need for a rapid identification technique. Enterococcus faecalis is readily identified by biochemical tests, but species differentiation within the Enterococcus faecium and Enterococcus gallinarum species groups is less well established. In the present study, 66 strains representing the most prevalent human enterococci were used to develop a PCR-based species-specific identification protocol. Whole-cell protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used as a reference method for species identification. In addition, the genomic SmaI macro-restriction fragment distribution of all of the strains was examined by pulsed-field gel electrophoresis (PFGE). Oligonucleotide D11344-primed PCR was as discriminative as whole-cell protein analysis and resulted in more easily interpreted band patterns. This PCR-based technique allowed identification of clinical isolates by visual examination of the DNA profiles obtained. The inability of both methods to discriminate between Enterococcus casseliflavus and Enterococcus flavescens brought into question the species status of E. flavescens. PFGE did not result in species-discriminative DNA bands or band patterns, but proved to be superior for interpretation of interstrain relationships.

Bacterial Typing Techniques↗
PubMed1997

Isolation of Candida species on media with and without added fluconazole reveals high variability in relative growth susceptibility phenotypes.

Mouthwashes from human immunodeficiency virus-positive individuals were sampled for yeasts by direct plating on a differential agar medium with and without added fluconazole and via enrichment broths with and without added fluconazole. The colonies of the yeasts isolated were tested for relative growth in the presence of single concentrations of itraconazole and fluconazole. Among 258 culture plates containing yeasts obtained via different isolation routes from 86 yeast-positive samples, 33 (12.7%) of the plates showed unexpectedly high colony-to-colony variation in relative growth. Intercolony variation was seen in 41 (47.7%) of the 86 isolates when relative growth data were analyzed for all colonies of an isolate tested, regardless of the medium used for isolation. The prevalence of relative growth variability with the azoles was highest for Candida glabrata (100% of 13 isolates), followed by Candida krusei (60% of 5 isolates) and Candida albicans (40% of 53 isolates), and the visual patterns of variability seen in scatter plots of the data showed species specificity. Relative growth phenotypes generally tended to be stable for each yeast colony in subcultures, whether or not the medium used for subculture contained antifungal agents. DNA fingerprinting of stable and variable C. albicans isolates showed changes in band patterns detected with the probe Ca3, suggesting that the variability may have resulted from selection of different subtypes of the yeasts during the isolation procedure. These findings suggest that the yeasts isolated from single clinical samples were often not clonal in nature. The relative growth test revealed colony variability more readily than conventional susceptibility testing.

Antifungal Agents↗
PubMed1997

Relevance of nucleic acid amplification techniques for diagnosis of respiratory tract infections in the clinical laboratory.

Clinical laboratories are increasingly receiving requests to perform nucleic acid amplification tests for the detection of a wide variety of infectious agents. In this paper, the efficiency of nucleic acid amplification techniques for the diagnosis of respiratory tract infections is reviewed. In general, these techniques should be applied only for the detection of microorganisms for which available diagnostic techniques are markedly insensitive or nonexistent or when turnaround times for existing tests (e.g., viral culture) are much longer than those expected with amplification. This is the case for rhinoviruses, coronaviruses, and hantaviruses causing a pulmonary syndrome, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, and Coxiella burnetii. For Legionella spp. and fungi, contamination originating from the environment is a limiting factor in interpretation of results, as is the difficulty in differentiating colonization and infection. Detection of these agents in urine or blood by amplification techniques remains to be evaluated. In the clinical setting, there is no need for molecular diagnostic tests for the diagnosis of Pneumocystis carinii. At present, amplification methods for Mycobacterium tuberculosis cannot replace the classical diagnostic techniques, due to their lack of sensitivity and the absence of specific internal controls for the detection of inhibitors of the reaction. Also, the results of interlaboratory comparisons are unsatisfactory. Furthermore, isolates are needed for susceptibility studies. Additional work remains to be done on sample preparation methods, comparison between different amplification methods, and analysis of results. The techniques can be useful for the rapid identification of M. tuberculosis in particular circumstances, as well as the rapid detection of most rifampin-resistant isolates. The introduction of diagnostic amplification techniques into a clinical laboratory implies a level of proficiency for excluding false-positive and false-negative results.

Bacterial Infections↗
PubMed1997

Comparison of screening methods for detection of extended-spectrum beta-lactamases and their prevalence among blood isolates of Escherichia coli and Klebsiella spp. in a Belgian teaching hospital.

Using a set of 33 well-defined extended-spectrum beta-lactamase (ESBL)-producing strains of Escherichia coli and Klebsiella pneumoniae, we compared three screening methods for ESBL detection: (i) a double-disk synergy test, (ii) a three-dimensional test (both the double-disk synergy test and the three-dimensional test were performed with ceftriaxone, ceftazidime, aztreonam, and cefepime), and (iii) the Etest ESBL screen (AB Biodisk, Solna, Sweden), based on the recognition of a reduction in the ceftazidime MIC in the presence of clavulanic acid. In the double-disk test, all four indicator antibiotics scored equally and 31 of the 33 reference strains were recognized. In the three-dimensional test, ceftriaxone was the only satisfactory indicator and 30 ESBL-positive strains were detected by this antibiotic. Both systems produced two false-positive results with cefepime. With the Etest ESBL screen, 15 of 16 TEM-related and 11 of 16 SHV-related ESBL-producing strains scored positive. In 10 cases the clavulanic acid on one end of the strip interfered with the MIC determination for ceftazidime, which was read on the opposite end. This MIC had to be determined with an extra ceftazidime-only strip. No false-positive results were noted. Eighty-six blood isolates of E. coli and Klebsiella species were screened for ESBL expression by the double-disk and three-dimensional tests, both with ceftriaxone. Six strains with suspicious antibiogram phenotypes also gave positive results by the double-disk test. One E. coli strain remained undetected by the three-dimensional test. Identification of the enzymes suspected of being ESBLs by isoelectric focusing (all strains) and DNA sequencing (1 strain) confirmed the screening test results except for one Klebsiella oxytoca strain, which proved to be a hyperproducer of its chromosomal enzyme and which also had a negative Etest score. The five true ESBL producers were all confirmed by the Etest ESBL screen. Pulsed-field gel electrophoresis proved that the E. coli strains were unrelated, but that two of the three K. pneumoniae strains were closely related.

Aztreonam↗
PubMed1997

New epidemiological features of meningococcal disease in Belgium.

SCIENTIFIC EDITOR In Belgium, after the epidemic of the early 1970s caused by Neisseria meningitidis B:2b:P1.2 (highest incidences in 1971 and 1972 with 5 cases per 100 000 population per year), the incidence of meningococcal disease fell to normal inter

Journal Article↗
PubMed1996

Typing of Mycoplasma pneumoniae by nucleic acid sequence-based amplification, NASBA.

Nucleic acid sequence-based amplification, NASBA, is an isothermal amplification technique for nucleic acids and was used for typing a collection of 24 Mycoplasma pneumoniae strains. A set of primers was chosen from the 16S rRNA sequence alignment of Mycoplasma species. The nucleotide sequences of the (-)RNA amplicons were determined for M. pneumoniae strains M15/83 (type 1) and FH (type 2), and revealed a one-point difference at the 16S rRNA level between the two types. Based on this result, two type-specific probes were constructed. The probes were hybridized in solution with the amplified nucleic acids of 24 M. pneumoniae strains in an enzyme-linked gel assay (ELGA). The results obtained by NASBA-based typing are in agreement with the classification of the 24 M. pneumoniae strains into two types by other typing methods, confirming the reliability of this technique.

Bacterial Typing Techniques↗
PubMed1996

In vitro activity of amoxycillin/clavulanate and ticarcillin/clavulanate compared with that of other antibiotics against anaerobic bacteria: comparison with the results of the 1987 survey.

The activity of amoxycillin/clavulanate (Augmentin) and ticarcillin/ clavulanate (Timentin) was tested against 351 strict anaerobic clinical isolates collected from September 1993 to April 1994 in eight Belgian university hospitals and compared with that of 8 other antibiotics using the NCCLS reference agar dilution procedure. Production of beta-lactamase was detected by the nitrocefin test in 48% of the isolates. At NCCLS-recommended breakpoints, more than 90% of isolates were susceptible to amoxycillin/clavulanate, ticarcillin/clavulanate, piperacillin/tazobactam, imipenem, chloramphenicol and metronidazole but only 77%, 72% and 48% to cefoxitin, clindamycin and penicillin, respectively. In comparison with the results of a similar survey conducted in 1987 no major changes in susceptibility were observed except for the susceptibility to clindamycin that declined from 83% to 72% overall, and from 83% to 66% in the B. fragilis group. Furthermore one isolate of Clostridium clostridioforme was found produce beta-lactamase and few B. fragilis group isolates showed reduced susceptibility to metronidazole.

Amoxicillin↗
PubMed1996

Evaluation of the Gen-Probe amplified Mycobacterium tuberculosis direct test for the routine diagnosis of pulmonary tuberculosis.

A total of 624 respiratory specimens from 543 patients (418 Belgian, 110 Rwandan, and 15 Colombian patients) were tested for the presence of Mycobacterium tuberculosis by the Mycobacterium Tuberculosis Direct Test (MTDT, Gen-Probe). Compared to culture, the MTDT on 497 samples of sputum or broncho-alveolar lavage from Belgium had a sensitivity, specificity and positive and negative predictive value of 86.4%, 96.0%, 50.0% and 99.3% respectively. The pooled results for Rwanda (112 specimens) and Colombia (15 specimens) were 97.8%, 65.7%, 88.2%, 92% respectively. After resolution of discrepant results by taking into account the clinical data, the results for the Belgian patients were 86.9%, 96.2%, 52.6%, 99.3% respectively, and for the Rwandan-Colombian patients 98.1%, 100%, 100% and 92% respectively. Results could be improved by testing more than one specimen from each patient and the inclusion of an internal control to detect inhibitors of the reaction. Culture remains necessary for drug susceptibility tests and the isolation and identification of non-tuberculous mycobacteria.

Belgium↗
PubMed1996

Severe infection due to Actinomyces bernardiae: case report.

We describe a patient with a serious urinary tract infection due to Actinomyces bernardiae (formerly classified as CDC [Centers for Disease Control and Prevention, Atlanta] fermentative coryneform group 2). Our patient had long-standing bladder dysfunction and developed hydroureteronephrosis, renal lithiasis, perirenal abscesses, and septicemia. A. bernardiae is distinct from Actinomyces pyogenes and several CDC coryneform groups.

Actinomyces↗
PubMed1996

Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of M. pneumoniae in acute respiratory tract infections in pediatric patients.

Mycoplasma pneumoniae and viruses in acute respiratory tract infections in children were studied during the winter of 1992-1993 in Antwerp, Belgium. M. pneumoniae was diagnosed in nasopharyngeal aspirates by culture and polymerase chain reaction (PCR). For this, amplification of a fragment of the PI adhesin gene in samples prepared by two methods was compared in two laboratories, and in one laboratory, a fragment of the 16S rRNA gene was amplified. The sensitivity of culture versus PCR was 61.5%. Provided a specific internal control is used, sample preparation by freeze-boiling combined with PCR for the PI gene and amplicon detection by visual inspection of the electrophoresis gel can be recommended, although maximal results are obtained after hybridization. M. pneumoniae was present in 0.5% of patients <2 years old and 6.9% of patients >2. M. pneumoniae was second to respiratory syncytial virus or detected equally in lower respiratory infections.

Adhesins, Bacterial↗
PubMed1996

Cefepime activity against Pseudomonas aeruginosa: evaluation of Etest and two disc diffusion methods.

The in-vitro susceptibility 244 consecutive, non-duplicate clinical isolates of Pseudomonas aeruginosa cefepime and ceftazidime was studied. Using an agar dilution method to measure susceptibility, the antibiotics showed comparable activity. The Etest and two types of agar diffusion discs were compared with the reference agar dilution method, and showed an absolute interpretive agreement value of over 95%.

Cefepime↗
PubMed1996

Polyphasic approach to the classification and identification of Gardnerella vaginalis and unidentified Gardnerella vaginalis-like coryneforms present in bacterial vaginosis.

A taxonomic study of Gardnerella vaginalis and G. vaginalis-like coryneforms was performed in order to clarify the phylogenetic affiliation of these organisms and to improve future identification. We examined 50 strains by performing whole-cell protein and fatty acid analyses, a 16S rRNA sequence analysis, and an extensive phenotypic characterization analysis. The results of both chemotaxonomic techniques which we used divided the organisms into two main clusters, and the 16S rRNA sequence analysis revealed that the clusters represent different genera, which were easily distinguished by the results of classical phenotypic tests. The cluster I strains were identified as G. vaginalis, which was shown to be a close relative of the genus Bifidobacterium. An improved description of G. vaginalis is presented. The cluster II strains belong to or are closely related to Actinomyces turicensis.

Bacterial Proteins↗