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See detailPrevalence and spread of extended-spectrum β-lactamase-producing
Magoue Lonchel, Carine ULg; MELIN, Pierrette ULg; Gangoué-Piéboji, J et al

in Clinical Microbiology & Infection (2013), 19(9), 416-20

During April 2010 and June 2010, 334 Enterobacteriaceae isolates from 590 participants (outpatients, inpatients, inpatient carers, hospital workers and members of their households) were collected from ... [more ▼]

During April 2010 and June 2010, 334 Enterobacteriaceae isolates from 590 participants (outpatients, inpatients, inpatient carers, hospital workers and members of their households) were collected from faecal samples. Based on b-lactamase pattern, origin of strains and the relationship between participants, 44 isolates of extended-spectrum b-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae were selected from 44 participants (in Ngaoundere Protestant Hospital and Ngaoundere Regional Hospital, Cameroon). To determine the relatedness of bacterial strains, these isolates were fingerprinted using the automated, repetitive-sequenced-based PCR-based DiversiLab system. Subsequently, E. coli isolates that had undergone DiversiLab analysis were examined with respect to their phylogenetic group and detection of the ST131 clone to shed light on the epidemiology of these isolates in the Ngaoundere hospitals. The prevalence of faecal carriage of ESBL-producing Enterobacteriaceae among the study participants was 54.06%. According to participant groups, the prevalence of faecal carriage was also high (outpatients 45%; inpatients 67%; inpatient carers 57%; hospital workers 44%; and members of their households 46%). Analysis of the molecular epidemiology of ESBL-producing E. coli and K. pneumoniae showed a close relationship of the isolates between related and nonrelated individuals. In addition, DiversiLab results of E. coli identified four related isolates (4/22) from cluster III belonging to the epidemiologically important clone ST131. Our results highlight the importance of outpatients, inpatients, their carers, hospital workers and their families as reservoirs of ESBLproducing Enterobacteriaceae. [less ▲]

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See detailGroup B streptococcal epidemiology and vaccine needs in developed countries
MELIN, Pierrette ULg; EFSTRATIOU, Androulla

in Vaccine (2013), 31(Supplement 4), 31-42

Development of a group B streptococcal vaccine (GBS) vaccine is the most promising approach for the prevention of GBS infections in babies, given the potential adverse effects of intrapartum antibiotic ... [more ▼]

Development of a group B streptococcal vaccine (GBS) vaccine is the most promising approach for the prevention of GBS infections in babies, given the potential adverse effects of intrapartum antibiotic prophylaxis as well as the need for effective prevention of both adult and late perinatal disease. There are numerous prevention strategies at this time but none are 100% effective in the eradication of neonatal early onset GBS disease and there are no preventative strategies for late onset disease. The need for a GBS vaccine is therefore, of utmost importance. Efforts applying genomics to GBS vaccine development have led to the identification of novel vaccine candidates. The publication of GBS whole genomes coupled with new technologies including multigenome screening and bioinformatics has also allowed researchers to overcome the serotype limitation of earlier vaccine preparations in the search of a universal effective vaccine against GBS. This review brings together the key arguments concerning the potential need of a GBS vaccine in developed countries and describes the current status with GBS epidemiology and microbiology in these countries. [less ▲]

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See detailPrevalence and mechanisms of resistance to carbapenems in Enterobacteriaceae
Huang, Te-Din; Berhin, Catherine; Glupczynski, Youri et al

in Journal of Antimicrobial Chemotherapy (2013), 68(8), 1832-7

Objectives: To determine the point prevalence of carbapenem-non-susceptible Enterobacteriaceae (CNSE) and carbapenemase-producing Enterobacteriaceae (CPE) isolates among hospitalized patients in Belgium ... [more ▼]

Objectives: To determine the point prevalence of carbapenem-non-susceptible Enterobacteriaceae (CNSE) and carbapenemase-producing Enterobacteriaceae (CPE) isolates among hospitalized patients in Belgium. Methods: Twenty-four hospital-based laboratories prospectively collected 200 non-duplicated Enterobacteriaceae isolates from clinical specimens of hospitalized patients over a 2 month period. All isolates were screened locally for decreased susceptibility to carbapenem drugs using a disc diffusion method according to CLSI interpretative criteria. CNSE strains were referred centrally for confirmation of carbapenemase by phenotypic and molecular testing. Results: From February to April 2012, 158 of the 4564 screened Enterobacteriaceae isolates were categorized as non-susceptible to carbapenems, resulting in a point prevalence of CNSE of 3.5% (95% CI: 2.9%–4.2%; range per centre: 0.5%–8.5%). Of the 125 referred CNSE isolates, 11 Klebsiella pneumoniae isolates [OXA-48 (n=7), KPC type (n=3) and NDM type (n=1)], 1 OXA-48-positive Escherichia coli isolate and 1 KPC-positive Klebsiella oxytoca isolate were detected in eight hospitals. None of the 72 carbapenem-non-susceptible Enterobacter spp. isolates were confirmed as CPE. The minimal estimated point prevalence of CPE isolates was 0.28% (13/ 4564; 95% CI: 0.13%–0.44%) overall (range per centre: 0%–1.5%). Conclusions: Despite the overall low prevalence of CNSE found in this study, the detection of CPE isolates in one-third of the participating centres raises concerns and highly suggests the spread and establishment of CPE in Belgian hospitals. [less ▲]

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See detailLe diagnostic microbiologique en 2013 ! Le parcours d'un échantillon.
MELIN, Pierrette ULg; DESCY, Julie ULg

Learning material (2013)

Ne vous êtes-vous jamais demandé à quoi pouvait ressembler un laboratoire de microbiologie clinique et quelles sont ses missions, de quelles façons les échantillons étaient traités pour obtenir la ... [more ▼]

Ne vous êtes-vous jamais demandé à quoi pouvait ressembler un laboratoire de microbiologie clinique et quelles sont ses missions, de quelles façons les échantillons étaient traités pour obtenir la détection et la caractérisation des agents pathogènes ? Le film présenté vous invite pour une visite virtuelle du secteur bactériologie du laboratoire de microbiologie médicale du CHU de Liège en suivant le parcours d’un échantillon de sa réception jusqu’au résultat final rendu au clinicien. Ce parcours comprend de multiples étapes en passant dans les mains de multiples personnes des plus expertes ainsi que la présentation des technologies analytiques et de communication utilisées au CHU de Liège. [less ▲]

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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 detailMicrobiological diagnosis of infectious keratitis
MELIN, Pierrette ULg

Conference (2013, March 16)

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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 detailExtended-spectrum β-lactamase-producing Enterobacteriaceae in Cameroonian hospitals
Lonchel, Carine Magoué; MELIN, Pierrette ULg; Gangoué-Piéboji, Joseph et al

in European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology (2013), 32(1), 79-87

Abstract Extended-spectrum β-lactamase (ESBL)-produc- ing Enterobacteriaceae have been described worldwide, but there are few reports on the carriage of these bacteria in Cameroon. In order to investigate ... [more ▼]

Abstract Extended-spectrum β-lactamase (ESBL)-produc- ing Enterobacteriaceae have been described worldwide, but there are few reports on the carriage of these bacteria in Cameroon. In order to investigate the types of ESBLs and to analyse some risk factors associated with ESBL carriage, faecal samples were collected between 3 January and 3 April 2009 from hospitalised patients at Yaounde Central Hospital and at two hospitals in Ngaoundere, Cameroon. Enterobacterial isolates resistant to third-generation cepha- losporins were screened for ESBL production using the double-disk synergy test. Polymerase chain reaction (PCR) and DNA sequencing were performed in order to find out the different types of ESBL genes in presumptive ESBL- positive isolates. During the study period, a total of 121 different patients were screened for ESBL carriage. The prevalence among these patients whose faecal samples were found to contain ESBL-producers was 55.3 % (67/121). According to a univariate analysis, hospitalisation during the previous year was found to be associated with ESBL carriage. Of the 71 bacteria isolated, Escherichia coli was predominant and represented 48 % of all isolates. ESBL characterisation revealed two types of ESBLs, CTX-M-15 (96 %) and SHV-12 (4 %). The present study emphasises the importance of screening for ESBLs in laboratories in Afri- can countries. The monitoring and detection of ESBL- producing bacteria are important in the setting up of appro- priate treatment of patients and to ensure effective infection control efforts. [less ▲]

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See detailBacteriological assessment of smoked game meat in Lubumbashi, D.R.C.
Kabwang a Mpalang, Rosette; Kakubu a Mpalang, Mireille; Mukeng Kaut, Clarence et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2013), 17

The bacteriological quality of smoked game meat in Lubumbashi has not been studied much to date. The present study focused on the analysis of 182 samples of smoked game meat from three species, Syncerus ... [more ▼]

The bacteriological quality of smoked game meat in Lubumbashi has not been studied much to date. The present study focused on the analysis of 182 samples of smoked game meat from three species, Syncerus caffer (n = 63), Phacochoerus aethiopicus (n = 60) and Sylvicapra grimmia (n = 59), sold at retail outlets in Lubumbashi. The isolation of Escherichia coli from 81.3% of samples (mean 4.87 ± 0.6 log10 CFU.g-1 of sample) confirms significant faecal contamination of smoked game meat. The study has determined by culture prevalences of 0.0%, 4.3% [CI95% 1.4-7.4], 3.8% [CI95% 1.1-6.6] and 14.2% [CI95% 9.2-19.4] respectively for Shiga toxigenic Escherichia coli (STEC), Salmonella spp., Campylobacter jejuni and Campylobacter coli. Using Polymerase Chain Reaction, these prevalences were of 2.2% [IC95% 0.1-4.3], 6.0% [IC95% 2.6-9.5], 3.8% [IC95% 1.1-6.6] and 15.9% [IC95% 10.6-21.3] respectively for STEC, Salmonella spp., C. jejuni and C. coli. Syncerus caffer was established as a potential vehicle of STEC carrying stx1 gene (3.2%), stx2 gene (1.6%) and the combination of stx2 and eae genes (1.6%). On the basis of these data, we suggested the need for developing monitoring plans of the production, preparation, handling and distribution of smoked game meat in Lubumbashi. [less ▲]

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See detailEUCAST Where are we Today? What’s New? A difficult road in Belgium.
MELIN, Pierrette ULg

Conference (2012, December 19)

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See detailCapsular Gene Typing of Streptococcus agalactiae Compared to Serotyping by Latex Agglutination
Yao, Kaihu; Poulsen, Knud; Maione, Domenico et al

in Journal of Clinical Microbiology (2012), 51(2), 503-507

We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalac- tiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping ... [more ▼]

We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalac- tiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping results obtained by latex agglutination. Among 281 human isolates 27% could not be typed by latex agglutination. All 312 isolates except 5 could be typed by the three PCR methods combined. Two of these methods were multiplex assays. Among the isolates that were typeable by both latex ag- glutination and capsular gene typing, 94% showed agreement between the two methods. However, each of the PCR methods showed limitations. One of the methods did not include all 10 recognized serotypes, one misidentified eight isolates of serotypes Ib and IV as serotype Ia, and one did not distinguish between serotypes VII and IX. For five isolates that showed aberrant pat- terns in the capsular gene typing, long-range PCR targeting the cps operon disclosed large insertions or deletions affecting the cps gene cluster. A sensitive flow cytometric assay based on serotype-specific antibodies applied to 76 selected isolates that were nontypeable by latex agglutination revealed that approximately one-half of these did express capsular polysaccharide. A proce- dure for convenient and reliable capsular gene typing to be included in epidemiological and surveillance studies of S. agalactiae is proposed. [less ▲]

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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 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 detailPERINATAL INFECTIONS - The GBS successful practices in prevention
MELIN, Pierrette ULg

Conference (2012, October 20)

Updated information on the most common infectious diseases that the mother may transmit to her infant during pregnancy, at birth or by breast feeding the infant. Most of the emphasis is directed to ... [more ▼]

Updated information on the most common infectious diseases that the mother may transmit to her infant during pregnancy, at birth or by breast feeding the infant. Most of the emphasis is directed to preventive measures, screening when interventions are available, and the detailed analysis of interventions during the preconceptional period, antenatal care, perinatal care and maternal and neonatal care after birth. The strategic approach for prevention is illustrated with the group B streptococci story. This information emphasizes the basic knowledge of the pathogen, the disease, the burden of problems caused by the disease in the mother and her offspring, epidemiological aspects, and how to manage the disease. Most of the emphasis is directed to screening-based preventive measures and new approaches to improve the accuracy and predictive values of screening are discussed. An update of the developments of a group B streptococcal vaccine is briefly presented. [less ▲]

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See detailEvaluation of a loop mediated isothermal amplification (LAMP) assay for the detection of group B streptococci
MELIN, Pierrette ULg

Conference (2012, October 09)

BACKGROUND GBS are still the leading cause of severe neonatal disease. Screening for vaginal-rectal GBS colonization in pregnant women at 35-37 weeks of gestation, followed by intrapartum ... [more ▼]

BACKGROUND GBS are still the leading cause of severe neonatal disease. Screening for vaginal-rectal GBS colonization in pregnant women at 35-37 weeks of gestation, followed by intrapartum antibioprophylaxis to colonized women has proven to be the most effective strategy for prevention of perinatal GBS disease. Despite GBS improved culture method, predictive values (PV) of prenatal screening for GBS colonization at delivery remain limited and contribute to ongoing disease. Switching from time-consuming cultures to a rapid molecular based assay for prenatal screening could be considered: a new assay, the illumigene® Group B Streptococcus (illuB) from Meridian Bioscience, based on LAMP technology is awaited to improve antenatal screening sensitivity. OBJECTIVE To evaluate the illuB performed on culture obtained by 18-24 hours incubation of vaginal-rectal swab in Lim broth versus the reference culture for GBS screening. To determine if and how the illuB may be integrated in the current recommended screening strategy. METHODS Vaginal-rectal specimens collected from 242 pregnant women, were sent to and processed by the labs of the University Hospital of Liege and Hospital Sint Lucas of Gent to determine the status of GBS colonization by the reference culture method for prenatal GBS screening and by illuB. Cultures were performed by direct plating onto Granada agar and inoculation in a selective enrichment Lim broth subsequently subcultured after overnight incubation onto Granada and StrepB Select agar. All illuB were performed from the same incubated Lim broth. Discrepant results were controlled by a PCR assay detecting a gene different from the target of the illuB. RESULTS GBS were recovered in culture from 20.7% of the specimens. By comparison with culture, sensitivity and specificity (S, Sp) of the illuB for identifying GBS colonization were 90% and 98.9%. The positive and negative PV (PPV,NPV) were 95.7% and 97.4% respectively. Among the discrepancies, the 2 illuB positive/culture negative results were also tested positive by the control PCR assay, confirming the presence of GBS DNA even if GBS were not cultured from these specimens. Overall for GBS detection, Sp and PPV were 100%. Out of the 5 false negative illuB, 3 matched cultures with very rare GBS meaning that GBS could have been absent in the aliquots used for illuB. Therefore, S and NPV of the illuB for the detection of GBS are probably higher than 90.4% and 97.4%. CONCLUSION The illuB demonstrated S and Sp close to those of the reference culture method and did not significantly improve the GBS detection. The higher cost of such molecular test may be an obstacle to its wide use as there is not a net benefice when compared with culture method; but a shorter turnaround time and limited hands on time could balance positively the decision. Additional clinical trials are needed. [less ▲]

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See detailGBS SCREENING Belgium: current and future guidelines
MELIN, Pierrette ULg

Conference (2012, October 02)

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See detailPhenotypical and Genotypical Surveillance of Macrolide and Lincosamide Resistance in Group B Streptococcus in Belgium
DESCY, Julie ULg; Ackermans, Yannick; BOREUX, Raphaël ULg et al

Poster (2012, September)

Background: Constant increase of erythromycin (E) and clindamycin (C) resistance (R) has been observed worldwide among isolates of group B streptococci (GBS). In Belgium, through the 2000s, E R increased ... [more ▼]

Background: Constant increase of erythromycin (E) and clindamycin (C) resistance (R) has been observed worldwide among isolates of group B streptococci (GBS). In Belgium, through the 2000s, E R increased rapidly from 10% to up to 30%. Therefore phenotypical and molecular surveillance of E and C R has to be conducted. Methods: 275 clinical isolates (N1) were obtained from a Belgian surveillance for invasive GBS disease in newborns (59 isolates with 32 early- and 27 late-onset diseases) and adults (216 strains) during 2008 to 2011 and 53 isolates (N2) from vagino-rectal colonization in pregnant women in 2010. E and C MICs were determined by using Etest® (EUCAST interpretive criteria). Furthermore, for the E R isolates, the inducible (iMLS), constitutive (cMLS) and M phenotypes were assessed by a double disk diffusion test; the distribution of genes encoding RNA methylases and efflux pumps was investigated by PCR. Results: Of the N1 and N2 isolates, 92 (33.5%) and 15 (28.3%) were respectively R to E, with a higher rate among serotype V (p <0.001) and serotype IV (p <0.05). Among these 107 E-R isolates, 100 (93.5%) exhibited the MLS phenotype (R to E and CC): 73 were cMLS with E MIC50 >256 mg/L and 27 iMLS with E MIC50/MIC90 12/>256 mg/L. The M phenotype (R to E and S to C) was expressed by 7 (6.5%) of E R isolates with E MIC50/MIC90 4/12 mg/L. One colonizing strain presented a newly described resistance mechanism in GBS: the L phenotype (S to E and R to C) with a C MIC at 8 mg/L. For cMLS, the most common E R genotype was ermB (66%) (p <0.05) followed by ermTR (29%) and ermB+ermTR (5%). All iMLS isolates harbored an ermTR gene except 3 (2 with ermB, 1 with both ermB and ermTR); and all M phenotype were positive for mefA/B gene. Conclusions:1) In Belgium, by year 2010, prevalence of macrolides R in GBS exceeded 30%, 2) MLS R phenotypes (target-site modification) were the majority mechanism; M phenotype (efflux R mechanism) was also prevalent. 3) E and C susceptibility testing and surveillance are mandatory to guide prophylaxis and treatment of serious GBS infections in penicillin-allergic patients (at high risk for anaphylaxis) but also to identify emergence of newly acquired resistance mechanisms such as the L phenotype. [less ▲]

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See detailPhenotypical and genotypical surveillance of macrolide and lincosamide resistance in group B streptococcus in Belgium
DESCY, Julie ULg; ACKERMANS, Yanick; BOREUX, Raphaël ULg et al

in Program and Abstract of the 52nd Intersciences Conference on Antimicrobial Agents and Chemotherapy (2012, September)

Background: Constant increase of erythromycin (E) and clindamycin (C) resistance (R) has been observed worldwide among isolates of group B streptococci (GBS). In Belgium, through the 2000s, E R increased ... [more ▼]

Background: Constant increase of erythromycin (E) and clindamycin (C) resistance (R) has been observed worldwide among isolates of group B streptococci (GBS). In Belgium, through the 2000s, E R increased rapidly from 10% to up to 30%. Therefore phenotypical and molecular surveillance of E and C R has to be conducted. Methods: 275 clinical isolates (N1) were obtained from a Belgian surveillance for invasive GBS disease in newborns (59 isolates with 32 early- and 27 late-onset diseases) and adults (216 strains) during 2008 to 2011 and 53 isolates (N2) from vagino-rectal colonization in pregnant women in 2010. E and C MICs were determined by using Etest® (EUCAST interpretive criteria). Furthermore, for the E R isolates, the inducible (iMLS), constitutive (cMLS) and M phenotypes were assessed by a double disk diffusion test; the distribution of genes encoding RNA methylases and efflux pumps was investigated by PCR. Results: Of the N1 and N2 isolates, 92 (33.5%) and 15 (28.3%) were respectively R to E, with a higher rate among serotype V (p <0.001) and serotype IV (p <0.05). Among these 107 E-R isolates, 100 (93.5%) exhibited the MLS phenotype (R to E and CC): 73 were cMLS with E MIC50 >256 mg/L and 27 iMLS with E MIC50/MIC90 12/>256 mg/L. The M phenotype (R to E and S to C) was expressed by 7 (6.5%) of E R isolates with E MIC50/MIC90 4/12 mg/L. One colonizing strain presented a newly described resistance mechanism in GBS: the L phenotype (S to E and R to C) with a C MIC at 8 mg/L. For cMLS, the most common E R genotype was ermB (66%) (p <0.05) followed by ermTR (29%) and ermB+ermTR (5%). All iMLS isolates harbored an ermTR gene except 3 (2 with ermB, 1 with both ermB and ermTR); and all M phenotype were positive for mefA/B gene. Conclusions:1) In Belgium, by year 2010, prevalence of macrolides R in GBS exceeded 30%, 2) MLS R phenotypes (target-site modification) were the majority mechanism; M phenotype (efflux R mechanism) was also prevalent. 3) E and C susceptibility testing and surveillance are mandatory to guide prophylaxis and treatment of serious GBS infections in penicillin-allergic patients (at high risk for anaphylaxis) but also to identify emergence of newly acquired resistance mechanisms such as the L phenotype. [less ▲]

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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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See detailGBS EPIDEMIOLOGY & VACCINE NEED IN DEVELOPED COUNTRIES
Melin, Pierrette ULg

Scientific conference (2012, July 06)

Development of a group B streptococcal vaccine (GBS) vaccine is the most promising approach for the prevention of GBS infections in babies, given the potential adverse effects of intrapartum antibiotic ... [more ▼]

Development of a group B streptococcal vaccine (GBS) vaccine is the most promising approach for the prevention of GBS infections in babies, given the potential adverse effects of intrapartum antibiotic prophylaxis as well as the need for effective prevention of both adult and late perinatal disease. There are numerous prevention strategies at this time but none are 100% effective in the eradication of neonatal early onset GBS disease and there are no preventative strategies for late onset disease. The need for a GBS vaccine is therefore, of utmost importance. Efforts applying genomics to GBS vaccine development have led to the identification of novel vaccine candidates. The publication of GBS whole genomes coupled with new technologies including multigenome screening and bioinformatics has also allowed researchers to overcome the serotype limitation of earlier vaccine preparations in the search of a universal effective vac- cine against GBS. This review brings together the key arguments concerning the potential need of a GBS vaccine in developed countries and describes the current status with GBS epidemiology and microbiology in these countries [less ▲]

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