References of "Huynen, Pascale"
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See detailLa Maladie de Lyme: aspects diagnostiques et prise en charge thérapeutique.
HUYNEN, Pascale ULg; GIOT, Jean-Baptiste ULg

Conference (2013, September 19)

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See detailSurveillance of serotypes and antimicrobial susceptibility profile in group B streptococcus (GBS) in Belgium
Melin, Pierrette ULg; SACHELI, Rosalie ULg; Sarlet, Gilles ULg et al

in Program and Abstract of the 53rd Intersciences Conference on Antimicrobial Agents and Chemotherapy. Washington, USA: ASM. (2013, September)

BACKGROUND Today GBS vaccines for prevention of severe neonatal disease through transplacental delivery of antibodies directly from immunized mothers are in advanced stage of development. For the ... [more ▼]

BACKGROUND Today GBS vaccines for prevention of severe neonatal disease through transplacental delivery of antibodies directly from immunized mothers are in advanced stage of development. For the introduction of any GBS vaccine there are urgent needs for pre and post vaccine enhanced surveillance studies of strains isolated from both neonatal diseases and vagino-rectal colonization of pregnant women. In Belgium, surveillance of invasive isolates is regularly done by the NRC. We report in this study a surveillance of colonizing isolates of GBS. METHODS In 2012, 344 GBS isolates were obtained from a Belgian surveillance for vagino-rectal colonization among pregnant women (max. 5 isolates/lab). Capsular types were determined by agglutination (Strep-B-latex, SSI, Denmark) and MICs by using a microdilution method (Sensititre) and Etest® (EUCAST interpretive criteria). Furthermore, for the erythromycin (E) resistant (R) isolates, the inducible (iMLS), constitutive (cMLS) and M phenotypes were assessed by a double-disk diffusion test. RESULTS Serotype III was the more common (27.6%) followed by V, II, Ia, Ib, IV, IX, VII and VI (18.1%, 16.4%, 13.4%, 7%, 4.7%, 2.5%, 0.8%, 0.5%) and 8.9% were non typable. All isolates were susceptible to penicillin ; 29% were R to E with a higher rate among serotypes IV and V (p<0.05). Among these E-R isolates, 93% exhibited the MLS phenotype (R to E and CC): 66% were cMLS with E MIC50>256 mg/L and 27% iMLS with E MIC50/MIC90 2/>8 mg/L. The M phenotype (R to E and S to C) was expressed by 7% of E-R isolates with E MIC50/MIC90 2/4 mg/L. CONCLUSION Compared with Belgian data relating to neonatal invasive strains (NRC reports) 1) Serotype V and II are more frequent and III less frequent among colonizing isolates 2) Prevalence of E-R is similar in percentage and phenotypes with the MLS R phenotype as major mechanism. Extended surveillance of both invasive and colonizing isolates is needed currently to prepare the follow-up in the future vaccine era. [less ▲]

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See detailImprovement of transport condition of swabs for group B streptococcal (GBS) screening
MELIN, Pierrette ULg; Dodémont, Magali; Sarlet, Gilles ULg et al

in Program and Abstract of the 53rd Intersciences Conference on Antimicrobial Agents and Chemotherapy. Washington, USA: ASM. (2013, September)

BACKGROUND For the screening-based strategy for prevention of perinatal GBS disease, CDC Guidelines as many others recommend use of appropriate transport media (Amies, Stuart, e.g.) and processing of ... [more ▼]

BACKGROUND For the screening-based strategy for prevention of perinatal GBS disease, CDC Guidelines as many others recommend use of appropriate transport media (Amies, Stuart, e.g.) and processing of specimen as soon as possible within 1 to 4 days. False negative cultures occur for several causes including lost of GBS viability during transport. Could Lim broth, recommended for the selective enrichment, and Granada tubes be used as transport media for swab? Simulating conditions of routine practice, Lim broth and Granada tubes, were evaluated in vitro as transport media. METHODS Tubes of 3 brands of Lim broth (Becton Dickinson, bioMérieux, Copan) and Granada tubes (bioMérieux) were inoculated with low inocula of 10-100 CFU of GBS. Each type of tubes was incubated at 4°C, room T° (RT) and 35°C. GBS were enumerated from each tube by subculture on blood agar after 1, 2, 3 and 4 days of storage at the different T°. All tests were processed in triplicates with 3 strains of GBS belonging to serotype Ia, III and V. RESULTS No difference of survival was observed between the 3 strains. T° had significant impact on GBS recovery for each type of tubes. At 4°C the viability was hardly sustained along the 4 days. At RT and 35°C, an increase >6 log of the inocula was observed. The increase of GBS density was sustained at least 4 days for the 3 brands of Lim broth. For the Granada broth, such increase was also observed but at day 3 for tubes incubated at 35°C, viability decreased and for some tubes, GBS subcultures were negative at day 3 or 4. CONCLUSION To improve sensitivity of GBS screening cultures, Lim broth could be recommended as a strong transport media and the advisable storage condition would be RT to 35°C up to 4 days. In this way, initiating selective enrichment culture at the time of collection of specimen would provide higher sensitivity even for low density of colonization. Transport at 4°C should be avoided in favour with RT to 35°C. Studies in clinical setting are expected. For Granada tubes, storage at RT was fine but improvement seemed restricted in time at 35°C as there was a loss of viability after 3 days. For Granada tubes, extended evaluation and delimitation of use are needed. [less ▲]

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See detailEvaluation of a new rapid test for the detection of norovirus antigen in comparison with Real Time RT-PCR
HUYNEN, Pascale ULg; Mauroy, Axel ULg; Gérard, Catherine ULg et al

Poster (2013, September)

Diagnosis of NoV infection mainly relies on molecular methods. A detection of viral antigens can also be performed by immunochromatographic assays, and may be useful in outbreak settings. The aim of this ... [more ▼]

Diagnosis of NoV infection mainly relies on molecular methods. A detection of viral antigens can also be performed by immunochromatographic assays, and may be useful in outbreak settings. The aim of this study was to compare the performances of the new RDT ImmunoCardSTAT!®Norovirus (Meridian Bioscience®, Europe) with a real time RT-PCR. [less ▲]

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See detailLes infections digestives et leur prévention (hors C.difficile)
HUYNEN, Pascale ULg

Scientific conference (2013, May 22)

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See detailLes norovirus: grands coupables méconnus de gastro-entérites.
HUYNEN, Pascale ULg

Conference (2013, May 04)

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See detailMéningite bactérienne: diagnostic microbiologique
HUYNEN, Pascale ULg

Scientific conference (2013, May 02)

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See detailComparison of Real-Time Aspergillus PCR with Platelia™AspergillusEIA in broncho-alveolar lavage fluids for the diagnosis of invasive aspergillosis in neutropenic and non-neutropenic patients
RUZICKA, NADIA; BOREUX, Raphaël ULg; LEVAUX, Laetitia ULg et al

Poster (2013, April 27)

Objectives. Because of low sensitivity of fungal cultures and lack of standardization of Aspergillus PCR, laboratory diagnosis of invasive aspergillosis still relies on galactomannan (GM) detection in ... [more ▼]

Objectives. Because of low sensitivity of fungal cultures and lack of standardization of Aspergillus PCR, laboratory diagnosis of invasive aspergillosis still relies on galactomannan (GM) detection in broncho-alveolar lavage (BAL) fluids and serum. The aim of this study was double: first, to assess the place of a 18S rRNA Aspergillus real-time PCR test performed in BAL fluid for the diagnosis of invasive aspergillosis (IA) in neutro- and non-neutropenic patients in comparison with GM detection; secondly, to evaluate the use of three different GM cut-off values. Materials and methods. A total of 111 neutropenic and non-neutropenic patients hospitalized at the University hospital of Liège from March to October 2012 with suspicion of IA were included in the study. A total of 138 broncho-alveolar lavage fluids were evaluated by three laboratory diagnostic methods: 1/ culture on Sabouraud agar slants with antibiotics (bioMérieux, France) incubated at 28°C for 28 days; 2/ GM detection (Platelia ™Aspergillus EIA, Biorad) using GM index cut-off values at 0.5, 0.8 and 1, performed three times a week; 3/ a real-time Aspergillus PCR assay performed daily and targeting the 18S rRNA genes by using an in-house method. Clinical, radiological and microbiological data were reviewed for classification of patients. Results. Nine patients developed probable or possible IA. The sensitivity/specificity/positive (VPP) and negative (NPV) predictive values (%) for culture, PCR, and GM using 0,5 as cut-off value were respectively 41/100/100/94, 58/97/70/96, and 91/83/34/99. The use of 0,8 and 1 as GM index cut-off values increased the specificity to 89 and 92% respectively, and the VPP to 44 and 54%. PCR had a better turn-around time and allowed the detection of Aspergillus colonisation. Conclusion: GM detection in BAL fluids using a cut-off value of 1 was the most efficient laboratory test for the diagnosis of IA in neutropenic and non-neutropenic patients. Despite a lower sensitivity, PCR had a better VPP, and allowed the detection of culture-negative Aspergillus colonisations. A shorter turnaround time (TAT) due to daily practice of PCR tests may reduce the time-to-treatment up to 24 hours. [less ▲]

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See detailLes infections respiratoires: méthodes de diagnostic au laboratoire.
HUYNEN, Pascale ULg

Conference (2013, February 08)

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See detailFATAL ALVEOLAR ECHINOCOCCOSIS OF THE LUMBAR SPINE
KEUTGENS, Aurore ULg; SIMONI, Paolo ULg; DETREMBLEUR, Nancy ULg et al

in Journal of Clinical Microbiology (2013), 51(2), 688-91

For the last ten years, the southern part of Belgium has been recognized as a low-risk endemic area for alveolar echinococcosis. This infection, caused by Echinococcus multilocularis, usually induces a ... [more ▼]

For the last ten years, the southern part of Belgium has been recognized as a low-risk endemic area for alveolar echinococcosis. This infection, caused by Echinococcus multilocularis, usually induces a severe liver condition, and can sometimes spread to other organs. However, alveolar echinococcosis involving bones has been described only very rarely. Here, a fatal case of spondylodiscitis due to E. multilocularis contracted in southern Belgium is reported. [less ▲]

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See detailVirologie et Syndromes Cliniques
HUYNEN, Pascale ULg

Scientific conference (2013, January 23)

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See detailVirologie et syndromes cliniques
HUYNEN, Pascale ULg

Learning material (2013)

Suite à leur entrée dans l’organisme, certains virus restent localisés au niveau de la porte d’entrée et sont responsables d’infections localisées. D’autres virus à l’inverse vont diffuser dans ... [more ▼]

Suite à leur entrée dans l’organisme, certains virus restent localisés au niveau de la porte d’entrée et sont responsables d’infections localisées. D’autres virus à l’inverse vont diffuser dans l’organisme, donnant lieu à des infections généralisées avec atteinte secondaire d’un ou plusieurs organes cibles. L’atteinte sélective de certains tissus ou organes définit le tropisme du virus. Par ailleurs, les déficits immunitaires favorisent la survenue de certaines affections virales qui présentent souvent, dans ce contexte, une sévérité particulière. On distingue principalement 8 grands syndromes résultant des infections virales : les dermatoses, les infections respiratoires, les gastro-entérites, les hépatites virales, les atteintes neurologiques, les infections fœtales et néonatales, les infections spécifiques d’organes et virus, et le SIDA. [less ▲]

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See detailLes norovirus : qui sont-ils ? Le point sur la question
HUYNEN, Pascale ULg

in NOSO-info (2013), XVII(1), 7-12

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See detailNorovirus: "new" enteropathogen agents
HUYNEN, Pascale ULg

Conference (2012, December 07)

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See detailClostridium difficile: méthodes de diagnostic microbiologique.
HUYNEN, Pascale ULg

Conference (2012, November 20)

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See detailDirect identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS: MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction.
MEEX, Cécile ULg; Neuville, Florence; DESCY, Julie ULg et al

in Journal of Medical Microbiology (2012), 61

In cases of bacteraemia, a rapid species identification of the causal agent directly from positive blood culture broths could assist clinicians in the timely targeting of empirical antimicrobial therapy ... [more ▼]

In cases of bacteraemia, a rapid species identification of the causal agent directly from positive blood culture broths could assist clinicians in the timely targeting of empirical antimicrobial therapy. For this purpose, we evaluated the direct identification of micro-organisms from BacT/ALERT (bioMérieux) anaerobic positive blood cultures without charcoal using the Microflex matrix-assisted laser desorption/ionization (MALDI) time of flight MS (Bruker), after bacterial extraction by using two different methods: the MALDI Sepsityper kit (Bruker) and an in-house saponin lysis method. Bruker's recommended criteria for identification were expanded in this study, with acceptance of the species identification when the first three results with the best matches with the MALDI Biotyper database were identical, whatever the scores were. In total, 107 monobacterial cultures and six polymicrobial cultures from 77 different patients were included in this study. Among monomicrobial cultures, we identified up to the species level 67 and 66 % of bacteria with the MALDI Sepsityper kit and the saponin method, respectively. There was no significant difference between the two extraction methods. The direct species identification was particularly inconclusive for Gram-positive bacteria, as only 58 and 52 % of them were identified to the species level with the MALDI Sepsityper kit and the saponin method, respectively. Results for Gram-negative bacilli were better, with 82.5 and 90 % of correct identification to the species level with the MALDI Sepsityper kit and the saponin method, respectively. No misidentifications were given by the direct procedures when compared with identifications provided by the conventional method. Concerning the six polymicrobial blood cultures, whatever the extraction method used, a correct direct identification was only provided for one of the isolated bacteria on solid medium in all cases. The analysis of the time-to-result demonstrated a reduction in the turnaround time for identification ranging from 1 h 06 min to 24 h 44 min, when performing the blood culture direct identification in comparison with the conventional method, whatever the extraction method. [less ▲]

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See detailSerological testing during pregnancy
HUYNEN, Pascale ULg

Conference (2012, October 12)

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See detailMolecular epidemiology of norovirus in symptomatic and asymptomatic population in Burkina Faso
HUYNEN, Pascale ULg; Mauroy, Axel ULg; Martin, Caroline et al

Poster (2012, September)

Background Noroviruses (NoV), belonging to the family Caliciviridae, are now recognized as the leading cause of gastroenteritis outbreaks worldwide, and represent an important cause of sporadic ... [more ▼]

Background Noroviruses (NoV), belonging to the family Caliciviridae, are now recognized as the leading cause of gastroenteritis outbreaks worldwide, and represent an important cause of sporadic gastroenteritis in both children and adults. Many studies describe NoV epidemiology. However, few data are available about the NoV strains circulating in most of African countries, in particular in Burkina Faso. The population of Burkina Faso is characterized by the young age of its habitants, and most are living in rural environment. Objectives The purpose of this epidemiological study was to determine the prevalence of NoV in Bobo Dioulasso (Southern part of Burkina Faso) by molecular diagnosis methods in patients presenting or not gastroenteritis symptoms, to quantify the excreted viral load, and to genotype the circulating strains. Methods Patients with and without gastro-intestinal disorders were selected in several Health Care Centres of Bobo Dioulasso. Clinical and epidemiological data, as well as stool samples, were collected during 8 weeks through March to April 2011. Viral genomic RNA was automatically extracted with a Maxwell® (Promega) instrument. Molecular detection of genogroups (G) I, II and IV NoV in stool samples was performed by a home-made real-time RT-PCR targeting the ORF1-ORF2 polymerase junction region. For each positive sample, viral load was estimated by using standard curves (successive dilutions of recombinant GI and GII plasmids). Molecular characterization was performed on the detected strains, using both polymerase and capsid regions. Results NoV were detected in 21.6% of the 453 collected stool samples, with a distribution of 21.0% and 23.1% in the samples from the 319 symptomatic (SP) and the 134 asymptomatic patients (AP) respectively. Genogroup distribution was 7.2% for GI, 10.7% for GII and 3.1% for both GI and GII among SP’s samples, and was 11.2% for GI, 10.4% for GII and 1.5% for both GI and GII among AP’s samples. Average viral load values were higher for GI NoV in SP than in AP (p=0.02), when they were higher for GII NoV in AP than in SP (p=0.04). Phylogenic analysis showed a high degree of genotypical diversity in both groups of patients. One recombinant strain GII.7/GII.6 was also detected, to our knowledge, for the first time. Conclusion Even if a true pathogenic role of NoV could not be showed from the study design, it allowed to precise the molecular epidemiology of NoV strains prevalent in a representative country of the East African region. It also showed that asymptomatic patients could play an important role as a NoV “reservoir”. Despite the fact that GII strains, and more precisely those belonging to GII.4 genotype, are nowadays highly reported worldwide, the surprising proportion of NoV GI detected in this study suggests that GI and GII strains should be excreted in equal proportion in the environment. The origin of this epidemiologic difference, even if partially explained by the difference in immunity and genetic sensitivity of the population, is still to be solved. [less ▲]

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