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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 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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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 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 detailSpectrométrie de masse MALDI-TOF en Microbiologie - 3ème BAC SBIM 2013
MEEX, Cécile ULg

Learning material (2013)

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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 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 detailAcute cholecystitis with Listeria monocytogenes
DESCY, Julie ULg; De Mol, Patrick ULg; HAYETTE, Marie-Pierre ULg et al

in Acta Clinica Belgica (2012), 67(4), 295-297

Listeriosis, an opportunistic food-borne disease caused by Listeria monocytogenes, is infrequent and occurs preferentially in patients at the extremes of age, during pregnancy or in immunocompromised ... [more ▼]

Listeriosis, an opportunistic food-borne disease caused by Listeria monocytogenes, is infrequent and occurs preferentially in patients at the extremes of age, during pregnancy or in immunocompromised hosts. Most common manifestations are maternofoetal and neonatal infections, severe invasive presentations such as bacteraemia with or without central nervous system symptoms occuring preferentially in immunosuppressed patients and self-limited gastro-enteritis affecting healthy individuals. Exceptionally, focal infections such as cholecystitis are described. We report here a case of acute cholecystitis caused by Listeria monocytogenes in an 82-year-old woman. Thanks to a successful treatment: cholecystectomy and antimicrobial therapy (amoxicillin plus clavulanic acid), the patient soon recovered. This case-report provides an opportunity to review the current literature concerning the association of Listeria monocytogenes and cholecystitis. [less ▲]

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See detailEvaluation of three immunoassays for serodiagnosis of human Mycoplasma pneumoniae infection
HUYNEN, Pascale ULg; TOUSSAINT, Françoise ULg; HAYETTE, Marie-Pierre ULg et al

in Clinical Microbiology & Infection (2012, April), 18(S3), 231

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See detailEvaluation of three immunoassays for serodiagnosis of human Mycoplasma pneumoniae infection
HUYNEN, Pascale ULg; TOUSSAINT, Françoise ULg; HAYETTE, Marie-Pierre ULg et al

Poster (2012, April)

The aim of this study was to evaluate three commercial automated immunoassays for the serological diagnosis of M. pneumoniae infection.

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See detailProportion of extended-spectrum ß-lactamase-producing Enterobacteriaceae in community setting in Ngaoundere, Cameroon
LONCHEL, Carine Magoué; MEEX, Cécile ULg; Gangoué-Piéboji, Joseph et al

in BMC Infectious Diseases (2012), 12

BACKGROUND: There is no information regarding the resistance mechanisms of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae in community setting in Cameroon. The current study aimed to ... [more ▼]

BACKGROUND: There is no information regarding the resistance mechanisms of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae in community setting in Cameroon. The current study aimed to determine the proportion of ESBLs in Enterobacteriaceae isolated in the community and to analyse some risk factors associated with ESBL carriage. METHODS: Faecal samples were collected from 208 different outpatients and 150 healthy student volunteers between 3 January and 3 April 2009. Enterobacterial isolates resistant to third-generation cephalosporins were screened for ESBL production by the double-disk synergy test. Presumptive ESBL-producing isolates with positive synergy test were identified by Mass Spectrometry using the BioTyper MALDI-TOF. For such ESBL positive isolates, antibiotic susceptibility was determined by the Vitek 2 system. PCR and sequencing were performed for the detection of different types of ESBL genes in presumptive ESBL-producing isolates. Statistical methods were used for the univariate calculation of risk factors. RESULTS: During the study period, a total of 358 faecal samples were analysed; 58 of such samples (16%) showed an ESBL phenotype and were confirmed by PCR. The proportion of ESBL producers in faecal carriage was statistically different between outpatients and student volunteers (23.1% vs. 6.7%: p < 0.000). According to a univariate analysis, previous use of antibiotics (ciprofloxacin) appeared to be a risk factor for ESBL carriage (p < 0.05).Escherichia coli was the species most frequently isolated among the ESBL producers in outpatients (66.7%) and student volunteers (90%). Isolates showed additional resistance to gentamicin, ciprofloxacin and trimethoprim/sulfamethoxazole but none of them was resistant to temocillin, amikacin or meropenem. Most of the strains (97%) produced a CTX-M group 1 enzymes [CTX-M-15 (98%) or CTX-M-1 (2%)] and the remaining strains produced SHV-12 enzyme (3%). CONCLUSIONS: The use of drugs such as amoxicillin, ciprofloxacin and trimethoprim/sulfamethoxazole does not seem appropriate for empirical treatment because of emerging resistance. The implementation in Cameroon or in other African countries of methods of screening ESBL-producing organisms in routine laboratories is of great importance in order for us to offer patients appropriate treatment and for infection control efforts to succeed. [less ▲]

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