Publications of Pascale HUYNEN
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See detailGroup B streptococcus neonatal invasive infections in Belgium 2010-2017, and characterization of isolated strains.
Melin, Pierrette ULiege; SACHELI, Rosalie ULiege; Lambotte, Olivia et al

in INMIS, International Committee (Ed.) INMIS 2019 Abstract book (2019, September)

Introduction/Background & Aims: Where intrapartum antibiotic-prophylaxis (IAP) is given to pregnant women colonized with Group B Streptococcus (GBS), the incidence of neonatal early-onset disease (EOD ... [more ▼]

Introduction/Background & Aims: Where intrapartum antibiotic-prophylaxis (IAP) is given to pregnant women colonized with Group B Streptococcus (GBS), the incidence of neonatal early-onset disease (EOD) has been successfully reduced; nevertheless, GBS is still the leading cause of severe disease among newborns, notably because the incidence of GBS late-onset disease (LOD) is not affected by IAP. Another strategy such as maternal immunization for prevention of both EOD/LOD is highly desirable worldwide. Aiming to describe GBS epidemiology and characterization of relevant epidemiological markers for vaccine development, surveillance of isolates causing neonatal disease is needed. We provide here results from the Belgian surveillance organized by the National Reference Centre(NRC). Methods: A total of 292 strains of GBS isolated from blood culture/cerebro-spinal fluid of newborns with invasive disease (149 EOD; 143 LOD) were sent to NRC by laboratories of a surveillance network, through years 2010-2017. Capsular-polysaccharide (CPS)-typing and pili-typing were performed with multiplex PCR assays. Multilocus sequence-typing and assignment to the hypervirulent clonal-complex (CC)17 was determined. Results: CPS type III isolates were responsible for 38.9% (n=58) of EOD cases, followed mainly by types Ia, V and II (22.1%, 18.1%, 8.1%). LOD cases were mainly caused by type III isolates (n=107, 74.8%), followed by types Ia (12.6%), V, Ib, IV and II (4.2%, 3.5%,2.8%, 2.1%). These distributions did not vary during the study period. A pili type was assigned to all isolates: at least one pili gene, PI2a, PI2b, or a combination of genes PI1-PI2a and PI1-PI2b. In 2016-2017, the hypervirulent-clone CC17 accounted for 33.3% of EOD (70.4% of type III) and 67% of LOD (89% of type III). Conclusions: The Belgian CPS distributions of isolates from EOD/LOD were similar to European data. One or 2 of 3 pilus-genes were detected in all isolates. CPS type III was predominant in both EOD/LOD and was mainly represented by CC-17 strains. [less ▲]

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See detailEvaluation of the Abbott RealTime quantitative CMV and EBV assays using the maxCycle protocol in a laboratory automation context
Bontems, Sébastien ULiege; BOREUX, Raphaël ULiege; CAPRARO, Valérie ULiege et al

in Journal of Virological Methods (2019), 270

Real-time PCR are often used for the diagnosis and monitoring of Cytomegalovirus (CMV)and Epstein-Barr virus (EBV)infections in susceptible populations. In this context, we evaluated the analytical ... [more ▼]

Real-time PCR are often used for the diagnosis and monitoring of Cytomegalovirus (CMV)and Epstein-Barr virus (EBV)infections in susceptible populations. In this context, we evaluated the analytical performances of the Abbott RealTime CMV/EBV maxCycle protocol automated on the m2000 platform (Abbott). It was compared to our routinely-used procedure consisting of a NucleoMag® DNA extraction automated on a STARlet platform followed by manually processed CMV and EBV quantitative real-time PCR (Diagenode). In this study, we showed that both EBV assays exhibited a similar sensitivity but with a better precision for the EBV Abbott RealTime assay. For the CMV performances, the Abbott assay was more sensitive and more precise than our routine method. The use of WHO International Standards also indicated a slight underestimation of the viral loads (−0.25 log10 IU/mL and −0.21 log10 IU/mL for CMV and EBV assays respectively)while these were rather overestimated with the Starlet/Diagenode method (0.48 log10 IU/mL and 0.19 log10 IU/mL for CMV and EBV assays respectively). These trends were confirmed using relevant whole-blood clinical samples and external quality controls. The workflows were also compared and we highlighted a significant technician hands-on time reduction (−63%)using the Abbott CMV/EBV maxCycle automated protocol. © 2019 Elsevier B.V. [less ▲]

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See detailDépistage intrapartum du Streptococcus agalactiae par PCR GenePOC GBS DS
MEEX, Cécile ULiege; Defêche, Justine ULiege; DEVEY, Anaïs ULiege et al

Poster (2018, December)

Les infections néonatales précoces causées par Streptococcus agalactiae (streptocoque du groupe B, GBS) peuvent être prévenues par administration d’une antibioprophylaxie intrapartum (AIP) aux femmes ... [more ▼]

Les infections néonatales précoces causées par Streptococcus agalactiae (streptocoque du groupe B, GBS) peuvent être prévenues par administration d’une antibioprophylaxie intrapartum (AIP) aux femmes identifiées GBS positives. Un test de dépistage vaginal rapide réalisé en début de travail permettrait une meilleure sélection des candidates à l’AIP que la stratégie basée sur un dépistage anténatal. OBJECTIFS Evaluation en laboratoire du test PCR GenePOCTM GBS DS (PCR GBS DS) sur frottis vaginal prélevé en intrapartum : performances et praticabilité du test en vue de sa réalisation en Point-Of-Care (POC). MATERIEL & METHODES De janvier à août 2018, inclusion de 102 frottis vaginaux prélevés en intrapartum avec le consentement de patientes admises en travail à la maternité du CHU de Liège. Collecte et évaluation se poursuivent. A la réception au laboratoire, décharge des frottis dans le milieu de conservation du kit PCR GBS DS: 150 µl utilisés pour réaliser le test PCR GBS DS sur système RevogeneTM, 10 µl mis en culture sur milieu sélectif Granada (Becton Dickinson, BD) et 300 µl inoculés en bouillon de Lim (BD). Après une nuit d’incubation, sous-culture sur Granada et gélose sélective chromogène StrepBselect (Biorad). Contrôle des discordances entre test PCR GBS DS et culture par PCR GenePOCTM GBS LB et PCR XpertTM GBS LB (Cepheid) sur les bouillons de Lim. RESULTATS Des cultures intrapartum, 12% étaient positives : 9 en primoculture et 3 après enrichissement. Le test PCR GBS DS a identifié 11 de ces échantillons positifs. Par comparaison avec la culture intrapartum, les sensibilité et spécificité du test PCR GBS DS, calculées sur base de ces premiers résultats, sont de 92 et 99% respectivement. Le taux d’erreur du test PCR GBS DS est de 2%. La réalisation du test PCR GBS DS et l’utilisation du système RevogeneTM sont extrêmement simples. Les résultats positifs ou négatifs sont obtenus en 75 minutes. DISCUSSION Les sensibilité et spécificité démontrées par le test PCR GBS DS réalisé au laboratoire sur frottis vaginaux intrapartum répondent aux exigences requises pour un dépistage intrapartum. Le délai de 75 minutes pour l’obtention des résultats dépasse les attentes idéales pour un test intrapartum. Ces performances devront être confirmées lorsque l’analyse sera réalisée en POC par des sages-femmes au bloc d’accouchement. [less ▲]

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See detailIl faut réintroduire le remboursement du dépistage du cytomégalovirus
HUYNEN, Pascale ULiege

Article for general public (2018)

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See detailVALIDATION, IMPLEMENTATION AND ROUTINE WORK WITH TOTAL LAB-AUTOMATION BD-KIESTRA : A TWO-YEAR EXPERIENCE AT THE UNIVERSITY HOSPITAL OF LIÈGE, BELGIUM
DESCY, Julie ULiege; MEEX, Cécile ULiege; BONTEMS, Sébastien ULiege et al

Poster (2018, April 21)

Background: Automation in Bacteriology is a major revolution in clinical microbiology laboratories. At the university hospital of Liège (Belgium), a BD-Kiestra Total Laboratory Automation (BD-K-TLA ... [more ▼]

Background: Automation in Bacteriology is a major revolution in clinical microbiology laboratories. At the university hospital of Liège (Belgium), a BD-Kiestra Total Laboratory Automation (BD-K-TLA) system has been installed in September 2015. Here, we describe the validation process achieved for the implementation of the BD-K-TLA system, and our two-year experience in the daily practice of the laboratory. Materials/methods: We present the validation plan (i.e. all the parameters that have to be checked before TLA-BD-K implementation) established by the Key User Group BD-Kiestra. In order to evaluate some of the added-value of this system compared to full-manual bacteriology, we measured two parameters for two similar period, before (February 2015) and after (February 2017) implementation of the BD-K-TLA: (1) the positivity rate (%), defined as the number of significant positive cultures among the total of samples analyzed and (2) the time-to-result (TTR), defined as the time (in hours) between reception of a specimen in the microbiology laboratory and the isolation of a significant pathogen in this sample. All specimens were included, except respiratory samples and blood cultures. Results: Validation plan included the checking process of each of the modules of BD-K-TLA: BarcodA, InoqulA, ProceedA, ReadA Compact and ReadA Browser. Furthermore, specimens were tested in parallel, on the BD-K-TLA and with full-manual bacteriological techniques: a minimum of 95% agreement was observed for qualitative and semi-quantitative assessment of Gram stain and cultures. The positivity rate went from 19,7% (pre-implementation – 1169 of 5941 samples analyzed), to 21,8% (post-implementation – 1391 of 6381 samples analyzed). Mean TTR for the specimens was of 26,95 h in February 2015, while it was 26,83 h in February 2017. Looking at these two measures, no significant time saving is currently observed. However, TLA-BD-K allows traceability through the whole process, quality of inoculation, fixed incubation times and better biological validation by clinical microbiologists thanks to remote pictures review. Conclusions: Working with BD-K-TLA allows a better quality and traceability of the bacteriological results, with a major benefit of remote pictures review. In order to observe time saving with BD-K-TLA, a well-considered configuration of the instrument and major reorganization of the workflow are mandatory. [less ▲]

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See detail2015 Surveillance of Group B Streptococcus strains isolated from invasive diseases among adults in Belgium : bacteriological and clinical characteristics.
SACHELI, Rosalie ULiege; MEEX, Cécile ULiege; DESCY, Julie ULiege et al

in Abstract book of 1st ISSAD 2018 (2018)

Background Even if Group B streptococcus (GBS) disease risk is highest during the first 3 months of life, GBS also causes significant morbidity and mortality among adults, especially in the elderly and ... [more ▼]

Background Even if Group B streptococcus (GBS) disease risk is highest during the first 3 months of life, GBS also causes significant morbidity and mortality among adults, especially in the elderly and immunocompromised or with chronic illnesses. We here provide an overview of bacteriological and clinical characteristics of GBS causing invasive diseases in non-pregnant adults in Belgium. Methods Overall 143 GBS strains isolated from invasive diseases among non-pregnant adults sent, on a voluntary-base, to the National Reference Centre (NRC) during the year 2015 by any laboratory located in Belgium were characterized: capsular polysaccharide (CPS)-typing by agglutination and/or with PCR, pili-typing with PCR, antimicrobial susceptibility testing, and detection of resistance genes with PCR. Results Among adult invasive isolates, CPS-type Ia was predominant (25.1%) followed by V, III, II, IV, Ib (23%, 19.6%, 9.8%, 9.8%, 7%) and VI, VII, VIII (1.4% each), IX (0.7%). One strain remained non typeable even with PCR. All strains were susceptible to penicillin. Rate of resistance to macrolides/lincosamides was 35.7%. ErmB, ErmTr and MefA genes were detected respectively within 45.2%, 27.4% and 19.5% among the resistant strains. One strain presenting the L profile (isolated resistance to clindamycin) harboured the LsaC gene. About pili-typing, the combined PI1, PI2a genes were predominant with 51.2% of the cases, followed by PI2a alone, the combined PI1, PI2b and PI2b alone (33.3%, 14.7%, 1.4%). The vast majority of strains were isolated from blood: 43% of bacteremia without reported focus, 19% originating from skin-and-soft tissue infections, 4.9% urosepsis, 4.2% bone-and-join infections, 2.8% endocarditis and 1.4% meningitis. Conclusion Among invasive adult diseases, GBS bacteriological characteristics were consistent with reported data among European countries. Macrolides/lincosamides resistance rate has slightly increased. Bacteremia without identified focus and skin-soft tissue infections were highly predominant; meningitis and endocarditis were less common but associated with serious morbidity and mortality. [less ▲]

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See detailCharacterization of Group B Streptococcus strains isolated from neonatal invasive diseases in Belgium, 2015.
SACHELI, Rosalie ULiege; MEEX, Cécile ULiege; DESCY, Julie ULiege et al

in abstract book of 1st ISSAD 2018 (2018)

Introduction Despite advances in preventive strategies, Group B Streptococcal (GBS) disease is still a leading cause of severe neonatal infections. The Belgian National Reference Centre (NRC) routinely ... [more ▼]

Introduction Despite advances in preventive strategies, Group B Streptococcal (GBS) disease is still a leading cause of severe neonatal infections. The Belgian National Reference Centre (NRC) routinely performs surveillances of GBS invasive strains. We here provide an overview of bacteriological characteristics of GBS causing invasive diseases in infants during the year 2015. Methods All GBS strains isolated from neonatal invasive diseases sent to the NRC during the year 2015 by any laboratory located in Belgium were characterized: capsular polysaccharide (CPS)-typing by agglutination and/or with PCR, pili-typing with PCR, antimicrobial susceptibility testing, and detection of resistance genes with PCR. Results A total of 44 GBS strains isolated from neonatal invasive diseases were available: 21 from Early Onset Diseases (EOD), 23 from Late Onset Diseases (LOD). Considering the incidence of GBS infections in Belgium, this collection represents for 2015 about 40% of GBS invasive diseases. Overall, CPS-type III was predominant (54.6%) followed by Ia, Ib, II, V, IV and VI (15.9%, 11.4%, 9.1%, 4.5%, 2.3%, 2.3%). All strains were susceptible to penicillin. Resistance to macrolides and lincosamides was showed in 22.7% of the strains and mainly linked to the presence of Erm genes: ErmB gene alone was expressed in 70% of the strains (n=7), one strain harboured the combination ErmB with MefA genes and another the ErmB with LsaC genes. One resistant strain did not express any of these four genes. About pili-typing, all strains harboured one of the PI-2 variants alone or in combination: the predominant type was PI1, PI2b (36.3%) followed by PI1, PI2a (27.7%), PI2a (25%) and PI2b (11.4%). Conclusion The ratio of EOD/LOD described in Belgium in 2015 remains quite stable since a few years. CPS-type and pili-type distributions, and resistance rate to macrolides/lincosamides are quite similar to European and North American observations done during the last decade. [less ▲]

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See detailGroup B Streptococcus neonatal invasive infections in Belgium, 2013-2016
MELIN, Pierrette ULiege; LECOMTE, Laurie; SACHELI, Rosalie ULiege et al

in Abstract book of 1st ISSAD 2018 (2018)

Background Since 2003, intrapartum antimicrobialprophylaxis (IAP) based on a universal screening for group B streptococcus (GBS) colonization at 35-37 weeks’ gestation is recommended in Belgium. Following ... [more ▼]

Background Since 2003, intrapartum antimicrobialprophylaxis (IAP) based on a universal screening for group B streptococcus (GBS) colonization at 35-37 weeks’ gestation is recommended in Belgium. Following, a decrease in the incidence of early-onset disease (EOD) was observed whereas late onset disease (LOD) have not changed. We describe the clinical and bacteriological characteristics of GBS neonatal invasive infections reported to the Belgian National Reference Centre (NRC) for GBS from 2013 to mid 2016. Methods On a voluntary base, all laboratories located in Belgium are invited to notify GBS neonatal invasive disease to the NRC, to send the isolate and fill a case-report questionnaire. This surveillance includes cases from 01.2013 to 07.2016. Following confirmation of identification, capsular-typing was performed by both agglutination and with PCR. Results Data from 157 GBS invasive neonatal cases were analysed: according to age at onset there were 70 EOD (median: day 0) and 57 LOD (median: 28 days). Among EOD cases, a male/female ratio was 0.75. The major types were III (35.7%), followed by Ia, II, V, Ib, IV, VI (20%, 14.3%, 12.9%, 10%, <10%). Fever and respiratory distress were frequently reported at onset. Meningitis was notified for 10,7% of cases. Among LOD cases, male/female ratio was 0.86. The predominant type was III (71.6%), followed mainly by Ia (13.4%). Gestational age at birth was significantly lower for LO cases and twins more frequent. The predominant characteristic at onset was fever and 32.5% developed meningitis. One death was reported in both groups of EOD and LOD. Antenatal GBS screening results and IAP were also analysed in regard to age at onset of diseases. Conclusions Clinical presentations were associated with age at onset of infection. Serotype III predominated in neonatal infections. Positive antenatal screening and appropriate IAP were not always protective for EOD and of course not for LOD. [less ▲]

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See detailA case of giant cell arteritis associated with culture-proven Coxiella burnetii aortitis
de Worm, S.; GIOT, Jean-Baptiste ULiege; Courtoy, C. et al

in International Journal of Infectious Diseases (2018), 69

A case of proven Coxiella burnetii aortitis, possibly associated with giant cell arteritis (GCA), is reported. A 72-year-old man, who is a hunter, presented with weight loss, fever, jaw claudication, and ... [more ▼]

A case of proven Coxiella burnetii aortitis, possibly associated with giant cell arteritis (GCA), is reported. A 72-year-old man, who is a hunter, presented with weight loss, fever, jaw claudication, and hardened temporal arteries associated with a persistent inflammatory syndrome and arteritis of the whole aorta, including the brachiocephalic arteries, as seen on 18F-fluorodeoxyglucose positron emission tomography/computed tomography. The diagnosis of GCA was retained, and treatment with prednisolone was started. Given the aneurysm of the abdominal aorta, the patient underwent replacement of the abdominal aorta with an allograft. Histology showed intense chronic arteritis attributed to atherosclerosis with dissection. However, Coxiella burnetii infection was confirmed by serology and then by culture and molecular biology on the surgical specimen. A combination of hydroxychloroquine and doxycycline was added to tapered prednisolone and the outcome was favourable. © 2018 The Author(s) [less ▲]

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See detailEvaluation of the Xpert®GBS LB test (Cepheid) performed on antenatal screening LIM enrichment broth for detection of Streptococcus agalactiae or group B streptococcus, compared to the reference culture method.
MEEX, Cécile ULiege; DUPONT, Audrey; SACHELI, Rosalie ULiege et al

Conference (2017, October 19)

PCR, performed on LIM enrichment broth compared wit the reference method, that is subculture on selective differential agar performed from the same incubated Lim broth inoculated with a vagino-rectal swab ... [more ▼]

PCR, performed on LIM enrichment broth compared wit the reference method, that is subculture on selective differential agar performed from the same incubated Lim broth inoculated with a vagino-rectal swab collected at 35-37 weeks’ gestation. Material/methods: During an 8-months period in 2015-2016, series of consecutive vagino/rectal swabs collected for antenatal GBS screening (at the university hospital of Liege, Belgium) were plated first on Granada agar and then inoculated in selective enrichment LIM broth. The incubated broth was further sub-cultured on Granada and Biorad StrepBselect agars. Moreover, a sterile swab immersed in the same incubated broth was further analyzed by a real-time PCR targeting GBS using the Xpert®GBS LB test on the GeneXpert® system (Cepheid). Results: Among the 288 antenatal screenings included in the study, 48 (16.7%) were positive for GBS using the culture reference method and 51 Xpert®GBS LB test were positive (17.7%), includin the 48 samples positive in culture and 3 additional specimens for witch culture remained negative. Considering the enriched culture as the gold standard, the sensitivity and specificity of the Xpert®GBS LB test were 100% and 98.8% respectively. Conclusions: The Xpert®GBS LB test performed on incubated LIM broth is at least as efficient as selective enriched culture for antenatal screening of GBS. The turnaround-time and hands-on-ti are much shorter for the Xpert® GBS LB but it is more expensive than culture method, which may limit its use. [less ▲]

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See detailEvolution of rate and genotypes of resistance to macrolide/lincosamide among invasive Group B Streptococcus (GBS): Development of a multiplex PCR tool for simultaneous detection of ErmB, ErmTr, MefA and LsaC resistance genes.
SACHELI, Rosalie ULiege; DESCY, Julie ULiege; MEEX, Cécile ULiege et al

Poster (2017, October)

Methods: A multiplex-PCR, using a set of specifically designed or already described (Kataja, 1999; Malbruny, 2011) primers was developed and used to detect, in GBS, three genes for erythromyc resistance ... [more ▼]

Methods: A multiplex-PCR, using a set of specifically designed or already described (Kataja, 1999; Malbruny, 2011) primers was developed and used to detect, in GBS, three genes for erythromyc resistance, ermB, ermTR, mefA and one gene for clindamycin-resistance lsaC. AdhP gene amplification was used as control for GBS identification. All(219) GBS isolates from invasive infections in newborns and adults received by the Belgian National Reference Center for GBS in 2015, and control strains were tested for erythromycin/clindamycin susceptibility (disk-diffusion/broth- microdilution) and for detection of resistance genes. Results: PCR products demonstrated the expected respective sizes. The method has been validated successfully according to ISO15189 analytical requirements. Of the 219 isolates, 67(30,67%) w resistant to erythromycin and/or clindamycin: 44/67(65,78%) showed a constitutive-MLS phenotype and 10/67(14,9%) the inducible-MLS phenotype. Among the constitutive-MLS strains, 73% harboured ErmB gene, 13% ErmTR, 7% ErmB+mefA and 7% ermB together with LsaC gene. The inducible-MLS strains harboured mostly ErmTr gene (89%) and the others the ErmB gene. Among the 10/67(14,9%) GBS strains with an M-phenotype (isolated resistance to erythromycin), the MefA gene was exclusively detected. Among the 3(4,48%) strains showing an isolated resistance to clindamycin (L-phenotype), the LsaC gene was detected. Conclusion: The developed multiplex PCR is able to detect simultaneously four genes involved in MLS resistance in GBS. In 2015, 30,6% of the invasive GBS strains isolated in Belgium were resist to macrolides and/or lincosamides. The emergence of the L-phenotype in GBS described since 2010, justifies the relevance to also detect LsaC gene together with ErmB, ErmTr and MefA. [less ▲]

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See detailUpdate of the characteristics of Group B Streptococci (GBS) colonizing pregnant women in Belgium: capsular-type distribution, pili characterization, antimicrobial susceptibility profile and Multiple Locus Sequence Types.
SACHELI, Rosalie ULiege; MEEX, Cécile ULiege; DESCY, Julie ULiege et al

Poster (2017, October)

Aim: Improving knowledge and characterization of GBS strains colonizing pregnant women in Belgium. Methods: In 2013, collection of 387 strains of GBS from 80 laboratories participating in a national ... [more ▼]

Aim: Improving knowledge and characterization of GBS strains colonizing pregnant women in Belgium. Methods: In 2013, collection of 387 strains of GBS from 80 laboratories participating in a national survey among pregnant woman. For each strain, determination of capsular-polysaccharide type agglutination and PCR, of pili-type by PCR and of antimicrobial susceptibility by disk-diffusion, broth-microdilution and detection of resistant genes by PCR. For serotype III strains, determination sequence-type by Multiple-Locus Sequence-Typing (MLST). Results: Serotype III was the most prevalent (28.5%) followed by serotypes V, Ia, II, IV and Ib (20.4%, 19.9%, 17.8%, 7%, 5.4%). Serotypes VI, VII and IX were found each once. All strains remained susceptible to penicillin (MICs: 0.03-0.125 mg/L) and other beta-lactams tested; 28.7% were resistant to erythromycin and 26.7% to clindamycin. With regards to pili, all 387 strains harboured one the PI-2 variants alone or in combination and 70.3% contained PI-1. The 110 serotype III isolates were resolved into 18 STs. The most common were ST-17 (35.5%) followed by ST-19 (30%) and ST- ST-27, ST-23 (<=5%). Conclusion: Among GBS from colonized pregnant women in Belgium: capsular-type and pili distributions, and MLST profile among type III strains were quite similar to reported data from Europ and USA during the last decade. As showed in this study, penicillin remains the first line drug of choice. On the contrary, resistance rates against macrolides/lincosamide, has reached a plateau since a decade, but it is noteworthy to notify the emergence of strains with isolated resistance to clindamycine. [less ▲]

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See detailA 5-year survey of dermatophytes strains circulating in Belgium
SACHELI, Rosalie ULiege; DARFOUF, Rajae ULiege; ADJETEY BAHUN, Akolé ULiege et al

Poster (2017, October)

Objectives Dermatophytosis refers to superficial fungal infections of keratinized tissues caused by keratinophilic dermatophytes. They are the most common cause of superficial fungal infections worldwide ... [more ▼]

Objectives Dermatophytosis refers to superficial fungal infections of keratinized tissues caused by keratinophilic dermatophytes. They are the most common cause of superficial fungal infections worldwide. Epidemiological studies regarding dermatophytes infections have been conducted in several countries and differences in the incidence and in etiological agents have been reported. That is why national surveillance of circulating strains causing dermatophytosis is critical. The National Reference Center (NRC) for mycoses conducted a survey on dermatophytes strains circulating in Belgium from 2012 to 2016. The present study was performed to assess the profile of dermatophytosis and to identify the species involved. Methods The Belgian NRC for mycoses (Leuven and Liège) collected 14227 strains between January 2012 and December 2016. The strains were obtained from clinically suspected fungal infections of skin, hair and nails. Strains were collected from Belgian laboratories in order to confirm the fungal identification which was performed by microscopy and in case of doubt by ITS sequencing. Results Among the 14227 samples collected, 6248 were identified as dermatophytes (44%). Trichophyton rubrum was the most prevalent species accounting for 61,3% (n=3820) of the strains collected from all sources, followed by T. mentagrophytes complex (19,2%, n=1199) according to the ancient classification (including T. interdigitale, T. benhamiae and T. mentagrophytes). Other less prevalent species were also recorded: M. audouinii (n=507, 8,1%), M. canis (n=210, 3,3%), T. tonsurans (n= 140, 2,2%), T. violaceum (n=133, 2,1%), T. soudanense (n=125, 2%), M. praecox (n=60, 0,96%) and E. floccosum (n=19, 0,3%) for the main ones. Our data show the predominance of anthropophilic species causing tinea capitis especially M. audouinii responsible for 43,4% (n=303/716) of hair/scalp infection with an increasing number from 2014 to 2016. Trichophyton soudanense, rarely observed in Belgium in the past, is an emerging agent of tinea capitis particularly since 2013, accounting for 11,3% (n=81) of the cases during the 5-year study period. Zoophilic strains such as M. canis which were well represented in the past epidemiology of tinea capitis are decreasing accounting for only 8,8% (n=63) of hair/scalp infection. Finally, our data confirm the high prevalence of T. rubrum as the main etiologic agent of onychomycosis (78,1%, n=3094/3968) followed by T. mentagrophytes complex (18,8%, n=743/3968). Both latter strains were also responsible for the majority of skin infections as they were isolated respectively in 46,2% (n=693/1612) and 21,7% (n=348/1612) of skin samples. Conclusions The present epidemiological survey provides recent data on the prevalence of all dermatophytes circulating in Belgium. Analyzing such data is critical for the establishment of measures for prevention and control of dermatophytes infections. Our study confirms the predominance of T. rubrum followed by species from the ancient T. mentagrophytes complex (T. interdigitale + T. benhamiae) in the Belgian population. This survey highlights also the persistent predominance of M. audouinii and the emergence of T. soudanense as causative agents of tinea capitis. [less ▲]

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See detailLaboratory diagnosis of syphilis
HUYNEN, Pascale ULiege

Conference (2016, September 26)

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See detailToxoplasma gondii, cytomegalovirus, and Treponema pallidum infections in pregancy: what's new?
HUYNEN, Pascale ULiege; LAZZAROTTO, Tiziana; PEYRON, François

in Wiener Klinisches Magazin (2016), 1(3), 1-8

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See detailHow efficient and automated can be Serology and Stool Testing?
HUYNEN, Pascale ULiege

in Year (2015), 13(9), 2-4

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See detailEpidemiological aspects and genotypic characterization of T.violaceum strains collected during a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULiege; Dekkers, Charlotte; GRAIDE, Hélène ULiege et al

in Mycoses (2015, October), 58(Supplement 4), 189

Objectives The last two years, clinical cases of tinea capitis caused by Trichophyton violaceum (T. violaceum), have been identified 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 Trichophyton violaceum (T. violaceum), have been identified in Belgium. To better understand the emergence of this species in the population, the Belgian National Reference Center (NRC Liège) launched a one-year 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 and T. violaceum strains isolated from hair to the NRC with a form to fill in including epidemiological data. The fungal strains were identified by microscopy or ITS sequencing in case of doubtful identification. The genotypic analysis was performed by the DiversiLab® system (bioMérieux) for DNA fingerprinting and analysis. Epidemiological data were analyzed with the help of a biostatistician. Results Amongst the collected isolates, 23 strains were confirmed as T.violaceum (results concerning the 116 M. audouinii strains have already been reported). Analysis of the epidemiological characteristics of the infected population shows that the main age category concerns 0-4 year-old children (n=9, 39,1%) with a sex-ratio M/F of 1.875. Data concerning the geographic origin of the family were present in 82,6% of the cases and reveal that patients were mainly of Ethiopian origin (n=8, 57,9% of known cases). One patient was also from Burundi showing that T. violaceum strains probably circulate mainly in East Africa. The genotypic analysis led to the distinction of 2 variants of T. violaceum. The major group was composed of 17 strains which were mainly collected in the North of Belgium and included also the reference strain (18/23, 83,3%). The other group (6 strains) was close to the major group but the analysis of the spectral superposition showed some differences between these two groups, defining two distinct variants of T. violaceum in the Belgian population. This second variant was mainly recovered from South Belgium (5/6, 83,3%). No correlation could be made between the genotypic group and a particular ethnical origin as Ethiopian subjects were found in both groups. Conclusion The DiversiLab® system proved to be an efficient method to investigate the molecular epidemiology of dermatophytes infections as reported previously for M. audouinii. These results show that two distinct isolates co-exist in Belgium providing evidence of genetic heterogeneity and a possible spread of one genotypic variant in a restricted geographic area or the co-existence of two variants circulating in different African communities. However, no clear correlation could be established between the appartenance to a group and epidemiological factors, such as age or ethnical origin. [less ▲]

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See detailQuality and Innovation, key factors for laboratory evolution
HUYNEN, Pascale ULiege

in Clinical Chemistry and Laboratory Medicine (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 detailHow efficiency and automated can be serology and stool testing?
HUYNEN, Pascale ULiege

in Clinical Chemistry and Laboratory Medicine (2015), 53(S1), 161

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