References of "Huynen, Pascale"
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See detailQuality and Innovation, key factors for laboratory evolution
HUYNEN, Pascale ULg

Conference (2015, June 23)

Today the laboratory has to face many challenges: constant increase of number of tests to be run, private labs that competes in reaching a lower TAT, disease outbreak that arises without the possibility ... [more ▼]

Today the laboratory has to face many challenges: constant increase of number of tests to be run, private labs that competes in reaching a lower TAT, disease outbreak that arises without the possibility of human control, like the recent mumps outbreak, the need to provide fast results in case of emergency or for transplants, the request to keep high level of traceability of all results, the accreditation of the lab are just some examples. With the same number of operators, year after year, new clinical needs have to be satisfied in a timely manner, with efficiency and without compromise in quality. The solution for us has been, across several year, to look for innovation. Moving from Elisa to chemiluminescence and therefore from open systems to close and state of the art systems, it has allowed us to face with success all those challenges. The availability of more and more infectious disease markers on fully automated analyzers, with good level of performance, have let us to cope with all the changes that have happened across more than a decade. Indeed innovation and quality are fundamental to support properly the laboratory evolution that occurred since today and it is still occurring. Innovation in our laboratory it is also represented by the introduction of automatized tests not only on serum and plasma specimens, but also on CSF (for Lyme disease diagnosis) and on stool matrix. In 2013 in fact we have introduced, among the assays already tested in our laboratory, two assays performed on this matrix, the C. difficile Toxin A&B and GDH, due to the possibility offered by the LIAISON® systems to run all of them on the same serology platform, without crosscontamination. New markers will be available in the near future, and our laboratory will be always able to meet the next clinical needs. [less ▲]

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See detailNorovirus: ce que les praticiens de soins devraient savoir.
HUYNEN, Pascale ULg

Conference (2015, June 01)

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See detailEpidemiological aspects and genotypic characterization of strains of Microsporum audouinii isolated in the context of a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULg; Dekkers, Charlotte; DARFOUF, Rajae ULg et al

Poster (2015, May)

Objectives The last two years, clinical cases of tinea capitis caused by Microsporum audouinii (M. audouinii), have increased in Belgium. To better understand the emergence of this species in the ... [more ▼]

Objectives The last two years, clinical cases of tinea capitis caused by Microsporum audouinii (M. audouinii), have increased in Belgium. To better understand the emergence of this species in the population, the Belgian National Reference Center (NRC) for dermatophytes launched a national survey in 2013. Epidemiological aspects and genotypic characterization of the strains were included. Methods The study was conducted from March 2013 up to February 2014. All Belgian laboratories were asked to send M. audouinii strains isolated from hair to the NRC with a form to fill in including epidemiological informations. The fungal strains were identified by microscopy or ITS sequencing in case of doubt. The genotypic analysis was performed by the DiversiLab® system (bioMérieux) for DNA fingerprinting and analysis. Epidemiological informations were analyzed with the help of a biostatistician. Results Among the collected isolates, 117 strains have been currently confirmed as M. audouinii. Analysis of the epidemiological characteristics of the infected population shows that the main age category concerns 5-9 year-old children (64%, p< 0,0001) with a sex-ratio M/F of 1.97. Data concerning the geographic origin of the family have been obtained in only 33,6% of the cases. It reveals that strains have been mainly isolated from patients with a Belgian nationality (43,6%) suggesting bias in the data collection. The geographic origin of the remaining group includes several African countries such as Congo (20,61%), Guinea (12,8%) and Burundi (5,12%). The genotypic analysis led to the distinction of 6 genotypic variants of M. audouinii. One of these variants was exclusively recovered from South Belgium (11 strains). The major group was composed of 96 strains, well distributed in different Belgium locations. Two other groups of three strains each were close to the major group but the analysis of the spectral superposition showed some differences between these groups. The two last groups were clearly different from the major group but species identification was confirmed by ITS sequencing. Conclusion The results of the genomic analysis by Diversilab, show that several groups of M. audouinii isolates co-exist in Belgium providing evidence of genetic heterogeneity. However, no clear correlation could be established between the appartenance to a group and epidemiological factors, such as the age or ethnical origin. ________________________________________ [less ▲]

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See detailEpidemiological aspects and genotypic characterization of strains of Microsporum audouinii isolated in the context of a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULg; Géron, Bénédicte; Dekkers, Charlotte et al

Poster (2015, April 28)

Objectives The last two years, clinical cases of tinea capitis caused by Microsporum audouinii (M. audouinii), have increased in Belgium. To better understand the emergence of this species in the ... [more ▼]

Objectives The last two years, clinical cases of tinea capitis caused by Microsporum audouinii (M. audouinii), have increased in Belgium. To better understand the emergence of this species in the population, the Belgian National Reference Center (NRC) for dermatophytes launched a national survey in 2013. Epidemiological aspects and genotypic characterization of the strains were included. Methods The study was conducted from March 2013 up to February 2014. All Belgian laboratories were asked to send M. audouinii strains isolated from hair to the NRC with a form to fill in including epidemiological informations. The fungal strains were identified by microscopy or ITS sequencing in case of doubt. The genotypic analysis was performed by the DiversiLab® system (bioMérieux) for DNA fingerprinting and analysis. Epidemiological informations were analyzed with the help of a biostatistician. Results Among the collected isolates, 97 strains have been currently confirmed as M. audouinii. Preliminary analysis of the epidemiological characteristics of the infected population shows that the main age category concerns 5-9 year-old children (84%) with a sex-ratio M/F of 1.95. Data concerning the geographic origin of the family have been obtained in only 45.8% of the cases. It reveals that strains have been mainly isolated from patients with a Belgian nationality (77%) suggesting bias in the data collection. The geographic origin of the remaining group (23%) includes several African countries. The genotypic analysis led to the distinction of 3 genotypic variants of M. audouinii. One of these variants was exclusively recovered from South Belgium (11 strains). The major group was composed of 85 strains, well distributed in different Belgium locations. The last group contains only one strain but this strain was significantly different from the two other variants. Conclusion The automated typing DiversiLab® system proved to be an easy and efficient method to investigate the molecular epidemiology of dermatophytes infections. These preliminary results show that, through Belgium, several groups of isolates co-exist for M. audouinii providing evidence of genetic heterogeneity. At this time all epidemiological informations have not yet been assessed while 35 strains of M. audouinii remain to be analysed genotypically to give definitive conclusions. [less ▲]

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See detailDiagnostic microbiologique des infections respiratoires virales en Pédiatrie
HUYNEN, Pascale ULg

Conference (2015, March 05)

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See detailInfections sexuellement transmissibles (IST):
HUYNEN, Pascale ULg

Conference (2014, November 14)

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See detailA clinical lab experience with an automated HIV Antigen/Antibody (Ag/Ab) combined assay
HUYNEN, Pascale ULg; TOUSSAINT, Françoise ULg; GERARD, Christiane ULg et al

Poster (2014, May 11)

OBJECTIVES: To describe the diagnostic performance of a new fourth-generation HIV Ag/Ab chemiluminescent immunoassay, available on the new LIAISON® XL analyser, in a clinical setting. METHODS: Through ... [more ▼]

OBJECTIVES: To describe the diagnostic performance of a new fourth-generation HIV Ag/Ab chemiluminescent immunoassay, available on the new LIAISON® XL analyser, in a clinical setting. METHODS: Through February 2012-October 2013, 12,438 samples of serum, received at our laboratory for screening for HIV infection were routinely tested with LIAISON® XL Murex HIV Ab/Ag assay (HIV-XL), which employs HIV-1, HIV-1 group O, and HIV-2 antigens and anti-p24 monoclonal antibodies in two coupled reagent cartridges, providing information of the overall Ab/Ag reactivity and detail of the specific reactivity for anti-HIV/HIV p24 antigen. Each serum with positive result or with negative result displaying a value close to the cut-off were sent to the regional AIDS-Reference Laboratory (RefLab) to perform confirmatory assays (PCR, Immunoblot). A previous verification of the HIV-XL demonstrated 100% sensitivity with a challenge panel of hundred positive sera provided by the RefLab. Performed external quality control was from United-Kingdom National External Quality Assessment Service (NEQAS). RESULTS: Out of the clinical samples, 12,312 non-reactive samples (including 6 negative results displaying a value close to the cut-off further confirmed true HIV negative), 64 Ab HIV reactive samples (all confirmed HIV-1 positive by immunoblot), including 4 samples reactive also for Ag HIV (confirmed positive by Ag assay/PCR), 42 Ab HIV reactive samples tested negative by immunoblot, and 20 Ag HIV reactive samples tested negative by the kit used for the Ag p24 detection in our HIV Reference Lab, have been found. All the 43 NEQAS specimens tested, 16 reactive and 27 non-reactive, were correctly classified. These results, considered all together, provide a calculated positive predictive value of 57.5% with an estimated specificity of 99.5% (with 95% confidence interval of 99.36-99.62%), and a calculated negative predictive value of 100% with an estimated sensitivity of 100.0% (with 95% confidence interval of 95.49-100%). CONCLUSIONS: In our experience HIV-XL showed excellent performance associated to all the advantages of a fully automated/random access instrument. [less ▲]

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See detailEVALUATION OF THE RAPID DETECTION OF ST-17 AND ST-1 GROUP B STREPTOCOCCI USING A MICROFLEX MALDI-TOF MS (BRUKER)
MEEX, Cécile ULg; SACHELI, Rosalie ULg; DESCY, Julie ULg et al

Poster (2014, May)

Objectives Clearly associated to neonatal meningitis, Group B streptococci (GBS) classified as sequence type-17 (ST-17) are defined as the “highly virulent” clone amongst GBS. The aim of this study was to ... [more ▼]

Objectives Clearly associated to neonatal meningitis, Group B streptococci (GBS) classified as sequence type-17 (ST-17) are defined as the “highly virulent” clone amongst GBS. The aim of this study was to evaluate an easy and rapid method, recently described to detect ST-17 and ST-1 GBS, based on distinguishing peak-shifts present on the protein spectrum of these 2 sequence types, using a Microflex (Bruker) matrix-assisted laser desorption/ionization time of flight mass spectrometer (MALDI-TOF MS). Methods This study was performed on 67 multi locus sequence typed (MLST) GBS originated from the Belgian and Czech National Reference Centers, including 18 ST-17 and 16 ST-1. After culture on blood agar, an ethanol/formic acid extraction was performed on each strain. Each extract was spotted once on a target plate, overlaid with 1 µl alpha-cyano-4-hydroxycinnamic acid matrix and further analysed by a Microflex MALDI-TOF MS. One spectrum per isolate was recorded, 240 laser shots being recorded for each spectrum. The spectra were further analysed using a Bruker prototype software, and 2 logarithmic values, one for ST-17 and one for ST-1, calculated from the intensities of the present and absent peaks, were obtained for each strain. If >0, this value indicated the presence of the specific sequence type. In a second step, the test was repeated on each strain with discordant result when compared with MLST. Results Compared with MLST method, the first analysis of the strains gave poor results, leading to very low sensitivities (77.8% for ST-17 and 50% for ST-1) but rather good specificities (85.7% for ST-17 and 98.0% for ST-1). After repeating the analysis on the strains with discordant result, sensitivity, 100% and 93.8%, and specificity, 87.8% and 98.0%, for ST-17 and ST-1 respectively were highly improved. Conclusion Since ST-17 and ST-1 GBS both show distinguishing peak-shifts on their protein spectrum, as described by Lartigue et al., the distinction of these 2 sequence types is now possible by MALDI-TOF MS. To our knowledge, this study is the first describing this application on a Microflex MS using a software to classify the strains. The observed results are promising but, given to the variability of the logarithmic value given by the software, the need to perform several measures on a same strain seems to be essential. After optimization of the analysis procedure, this rapid, easy and cheap method could be used to precociously detect ST-17 among GBS isolated from prenatal screenings, allowing a better follow up of the colonized mothers and a closer monitoring of their newborns. We would like to thank the Bruker Company which allowed us to evaluate the prototype software they have developed. [less ▲]

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See detailRetour en force des oreillons chez les jeunes adultes. Faut-il un nouveau vaccin ?
HUYNEN, Pascale ULg

Conference (2014, April 04)

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See detailRetour en force des oreillons chez les jeunes adultes. Faut-il un nouveau vaccin ?
HUYNEN, Pascale ULg

Conference (2014, March 27)

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See detailMolecular epidemiology of norovirus infections in symptomatic and and asymptomatic children from Bobo Dioulasso, Burkina Faso
HUYNEN, Pascale ULg; Mauroy, Axel ULg; Martin, Caroline et al

in Journal of Clinical Virology (2013), 58

Background: Noroviruses (NoV) are a leading cause of gastroenteritis worldwide. Few epidemiologicaldata regarding the NoV strains circulating in African countries are available.Objectives: To determine ... [more ▼]

Background: Noroviruses (NoV) are a leading cause of gastroenteritis worldwide. Few epidemiologicaldata regarding the NoV strains circulating in African countries are available.Objectives: To determine the prevalence of NoV in Bobo Dioulasso (Burkina Faso) in both symptomaticand asymptomatic gastroenteritis patients.Study design: Patients both with and without gastro-intestinal disorders were selected. Clinical andepidemiological data, as well as stool samples, were collected through March to April 2011.NoV molecular detection (genogrouping and genotyping) and viral load quantification were also per-formed for all samples.Results: NoV were detected in 22.2% of the 418 collected stool samples (21.2% and 24.8% from the 293symptomatic patients (SP) and the 125 asymptomatic patients (ASP) respectively).Genogroup (G) distribution was 7.5%, 10.2% and 3.4% for GI, GII and both GI/GII respectively among SPand 12.0%, 11.2% and 1.6% for GI, GII and both GI/GII, respectively, among ASP.Average viral load values were higher in SP than in ASP for GI (p = 0.03) but not for GII.Phylogenic analysis showed a high degree of genotype diversity in SP and ASP. One recombinantGII.7/GII.6 sequence was, to the best of our knowledge, detected for the first time.Conclusions: This study enabled identification of the specific molecular epidemiology of NoV strains cir-culating in a representative country in Eastern Africa, and additionally showed that ASP could play animportant “reservoir” role. A high strain diversity was detected with a surprisingly high proportion ofNoV GI compared to the common genotypes usually reported in comparable epidemiological studies. [less ▲]

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See detailMaladies Sexuellement Transmissibles
HUYNEN, Pascale ULg; LIBOIS, Agnes

Scientific conference (2013, October 10)

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See detailDNA fingerprinting using Diversilab system for genotyping characterization of Microsporum audouinii and Trichophyton violaceum
SACHELI, Rosalie ULg; DIMO, Lauryl; GRAIDE, Hélène ULg et al

in Mycoses (2013, October 01), 56(Supplement S3), 99

Objectives: To investigate the epidemiological determinants responsible for the high number of anthropophilic dermatophytes received by the National Reference Center for Mycosis of Liege (NRCL) during the ... [more ▼]

Objectives: To investigate the epidemiological determinants responsible for the high number of anthropophilic dermatophytes received by the National Reference Center for Mycosis of Liege (NRCL) during the year 2012. To perform a genotypic characterization by the Diversilab® system focusing on the two main isolated species, Microsporum audouinii and Trichophyton violaceum. To present a preliminary study preceding the national survey launched in 2013. Methods: A total of 51 strains of M. audouinii (50 clinical + 1 reference (ref.) strains) and 15 strains of T. violaceum (14 clinical + 1 ref. strain) originating from different locations through Belgium were included in the study. The fungal strains were first cultivated on Malt agar, then sub-cultured in Sabouraud liquid medium (Fluka). The grown mycelium was processed for DNA extraction following recommendations of the manufacturer (Ultra Clean® DNA Microbial isolation kit, MoBio laboratories). Genotypic analysis was performed using the DiversiLab® system (BioMérieux) for DNA fingerprinting and analysis. Results: Regarding M. audouinii, four different genotypic groups of strains were separated. Group 1 includes 11 strains and is only found in the Liège surroundings. Group 2 includes only one strain with little differences compared to group 1 and collected from the Liège area. These two groups may be related to each other. Group 3 contains 36 strains and the reference strain. This genotype is distributed in different Belgium locations. The last group, group 4, contains only 3 isolates sharing low similarities in comparison with the 3 other groups. Concerning T. violaceum, 6 different genotypic groups with a mixed geographical distribution were determined. Group 1 includes 8 clinical isolates and the ref. strain. The other five isolates are all different and seem not to be related to each other. Conclusion: The automated typing DiversiLab® system proved to be an easy and efficient method to investigate the molecular epidemiology of dermatophytes infections. Preliminary results of the study show that, through Belgium, several groups of isolates co-exist for M. audouinii and T. violaceum providing evidence of genetic heterogeneity. This variation can be related to acquired mutations due to environmental adaptation. Further investigations are necessary to better understand the impact of this genotypic variation. [less ▲]

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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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