References of "Hayette, Marie-Pierre"
     in
Bookmark and Share    
Full Text
Peer Reviewed
See detailInvestigation on possible transmission of monkeys' Plasmodium to human in a populations living in the equatorial rainforest of the Democratic republic of Congo
MVUMBI, Dieudonné; BOBANGA, Thierry; UMESUMBU, Solange et al

in International Journal for Parasitology: parasites and wild life (2016), 5

Plasmodiums are protozoa that may infect various hosts. Only five species are now recognized as naturally parasitizing humans: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ... [more ▼]

Plasmodiums are protozoa that may infect various hosts. Only five species are now recognized as naturally parasitizing humans: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi. This fifth species, P. knowlesi, previously identified as naturally parasitizing the monkey Macaca fascicularis, has been microscopically confused for a long time with P. malariae or P. falciparum and it was not possible to correctly differentiate them until the advent of molecular biology. To date, natural human infections with P. knowlesi only occur in Southeast Asia and a similar phenomenon of natural transmission of simian plasmodium to humans has not been reported elsewhere. This study was conducted to investigate a possible transmission of African small monkey's plasmodium to humans in populations living near the rainforest of the Democratic Republic of Congo (DRC) where several species of non-human primates are living. Two successive real-time PCRs were identified in the literature and used in combination for purpose. Only P. falciparum was found in this study. However, studies with larger samples and with more advanced techniques should be conducted. [less ▲]

Detailed reference viewed: 26 (5 ULg)
Full Text
See detailNew technologies in Mycology: PCR for diagnosis
Hayette, Marie-Pierre ULg

Conference (2015, November 19)

Molecular biology techniques are widely used in mycology. First of all they are used for confirmation of difficult or uncertain identifications. In this case PCR followed by ITS sequencing is generally ... [more ▼]

Molecular biology techniques are widely used in mycology. First of all they are used for confirmation of difficult or uncertain identifications. In this case PCR followed by ITS sequencing is generally used. For diagnostic purposes, PCR is mostly applied for the detection of invasive infections such as pneumocystosis, candidosis or aspergillosis. However these techniques (Candida and Aspergillus detection) are still not included in the recommandations given by EORTC by lack of standardisation. However, the development of new commercially available techniques, will probably change the diagnostic algorithm. New commercial PCR kits applied to the detection of dermatophytes are also recently available to the laboratories and they will be a good complement to culture-based diagnostic methods. [less ▲]

Detailed reference viewed: 21 (3 ULg)
Full Text
See detailFungal skin infections: from the clinic to the laboratory
Hayette, Marie-Pierre ULg

Conference (2015, November 14)

Superficial fungal infections are largely represented in the world population and are mainly caused by dermatophytes. Clinical signs can be confusing and it is important to get a rapid diagnostic to apply ... [more ▼]

Superficial fungal infections are largely represented in the world population and are mainly caused by dermatophytes. Clinical signs can be confusing and it is important to get a rapid diagnostic to apply the right treatment.The diagnostic procedures are still culture-based methods but molecular technologies are recently available under commercial kits and will be probable part of a new algorithm in the laboratory diagnosis. [less ▲]

Detailed reference viewed: 21 (0 ULg)
Full Text
Peer Reviewed
See detailValidation of the DermaGenius Nail plus multiplex assay, a new commercial PCR assay developed for the detection and identification of dermatophyte and Candida in nails
Hayette, Marie-Pierre ULg; GRAIDE, Hélène ULg; ADJETEY BAHUN, Akolé ULg et al

in Mycoses (2015, October), 58(suppl 4), 223

Objectives Superficial dermatophytosis is the most common fungal infection in humans. Diagnosis of dermatophytosis is currently based on microscopy or histology associated with culture on specific agar ... [more ▼]

Objectives Superficial dermatophytosis is the most common fungal infection in humans. Diagnosis of dermatophytosis is currently based on microscopy or histology associated with culture on specific agar media. However, direct microscopy lacks specificity and culturing has a long turn-around-time of 2-4 weeks. These limitations can be prevented by the use of molecular diagnostics. The DermaGenius (DG) multiplex kit (PathoNostics, The Netherlands) is a new commercial realtime-PCR kit, which can differentiate various dermatophytes species including the nail pathogens T. rubrum, T. mentagrophytes, T. interdigitale and 2 Candida species (C. albicans and C. parapsilosis). This study aimed at the validation of the kit on nails clippings. Results were compared with histology and culture data. Methods A set of 76 nail clippings was collected from 76 patients attending the dermatology consultation at the University Hospital of Liege on suspicion of onychomycosis. All nails were divided in three pieces for histology, culture and the PCR multiplex assay. Histologic preparations were stained with PAS staining. Cultures were performed on 2 different Sabouraud agar medium slants (bioMérieux). The DNA extraction protocol used a proteinase K pre-treatment followed by an automated DNA-extraction (EasyMag, bioMérieux). An Internal Control (IC) was included to monitor for PCR inhibition or manual errors. The realtime-PCR amplification was performed with the DG kit on a Rotor-Gene Q instrument (Qiagen) by using quantitative amplification and melting curve analysis. Results In total, 35 of 76 cases (46%) were classified as confirmed onychomycosis based on positive microscopy (M+) with or without positive culture (C+) or just by positive culture of a confirmed pathogen. Based on negative microscopy (M-) and negative culture of a confirmed pathogen, 41 cases (54%) were reported as non-fungal onychodystrophy. Agreement between DermaGenius (DG) and culture was found in 52% of the cases while 86% agreement was reported when comparing positive DG with confirmed onychomycosis. Three positive cultures (microscopy negative) were not detected by DG (2 T. rubrum, 1 C. albicans). However, DG could detect 7 additional infections (9%). Eleven discrepancies DG+/C+ were determined which could be positively confirmed in favour of DG result by ITS sequence analysis. Most discrepancies could be explained by fungal/yeast species overgrowing the agar slant, including species of Candida, Fusarium, Trichosporon, which were not considered as the source of infection. Conclusion The DermaGenius Nail plus multiplex was able to detect the most prevalent pathogenic dermatophytes species in clinical nail specimens and proved to be more sensitive and specific than culture and direct microscopy. The DNA extraction procedure has been shown to work efficiently in diagnostics which enables the physician in charge of the patient to start a dedicated treatment rapidly. [less ▲]

Detailed reference viewed: 42 (0 ULg)
Full Text
Peer Reviewed
See detailFalciparum malaria molecular drug resistance in the Democratic Republic of Congo: a systematic review
Mvumbi, Dieudonné; Kayembe, Jean-Marie; Situakibanza, Hippolyte et al

in Malaria Journal (2015), 14

Background: Malaria cases were estimated to 207 million in 2013. One of the problems of malaria control is the emergence and spread of Plasmodium falciparum strains that become resistant to almost all ... [more ▼]

Background: Malaria cases were estimated to 207 million in 2013. One of the problems of malaria control is the emergence and spread of Plasmodium falciparum strains that become resistant to almost all drugs available. Monitoring drug resistance is essential for early detection and subsequent prevention of the spread of drug resistance by timely changes of treatment policy. This review was performed to gather all data available on P. falciparum molecular resistance in DR Congo, as baseline for future assessments. Methods: The search for this review was undertaken using the electronic databases PubMed and Google Scholar using the terms “malaria”, “Congo”, “resistance”, “molecular”, “antimalarial”, “efficacy”. Articles were classified based on year of collecting, year of publication, sample size and characteristics, molecular markers analysed and polymorphisms detected. Results: Thirteen articles were included and five genes have been analysed in these studies: pfcrt, pfdhps, pfdhfr, pfmdr1 and K13-propeller. The majority of studies included were not representative of the whole country. Conclusion: This systematic review demonstrates the lack of molecular resistance studies in DRC. Only 13 studies were identified in almost 15 years. The MOH must implement a national surveillance system for monitoring malaria drug resistance and this surveillance should be conducted frequently and country-representative. [less ▲]

Detailed reference viewed: 23 (2 ULg)
Full Text
Peer Reviewed
See detailA three year survey of dermatophytoses in Belgium
SACHELI, Rosalie ULg; DARFOUF, Rajae ULg; GRAIDE, Hélène ULg et al

in Mycoses (2015, October), 58(Supplement 4), 135

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 dermatophyte infections have been conducted in several countries and differences in the incidence and in etiological agents have been reported for different geographical areas. That is why national surveillance of circulating strains causing dermatophytosis is crucial. The Belgian National Reference Center (NRC) for Mycoses conducted a survey on dermatophytes strains circulating from 2012 to 2014. The present study was performed to assess the profile of dermatophytosis and to identify the species involved. Methods The Belgian NRC for Mycosis collected 9138 strains between January 2012 and December 2014. The isolates were cultured from patients clinically suspected for fungal infections of skin, hair and nails. Isolates were sent by Belgian laboratories to the two labs of the Belgian NRC (UZ Leuven and CHU of Liège) in order to identify the fungus or to confirm the identification. All isolates cultured from patients of UZ Leuven and CHU of Liège were also included. Fungal identification was performed by microscopy after subculture and in case of doubtful identifications by ITS sequencing. Results .Among the 9138 samples (results of UZ Leuven and CHU of Liège combined), 3587 were identified as dermatophytes. Trichophyton. rubrum (T. rubrum) was the most prevalent species accounting for 56,17% (n=2015) of the infections from all sources, followed by T. mentagrophytes complex (21,83%, n=783). The other main etiological agents of dermatophytosis recorded in this study in descending order of prevalence were M. audouinii (n=303), M. canis (n=120), T. violaceum (n=112), T. tonsurans (n= 95), T. soudanense (n=66), M. praecox (n=59), E. floccosum (n=14) Our data reveal the predominance of anthropophilic species causing tinea capitis especially M. audouinii responsible for 36,49% (n=163/448) of hair/scalp infection. Trichophyton violaceum rarely observed in our country is frequently found as 12,8% (n=57) of the reported cases of tinea capitis are due to this species. The retrospective evaluation of data collected also shows that zoophilic strains as M. canis well represented in the past epidemiology of tinea capitis, is decreasing in frequency accounting for only 7,2% (n=32) of clinical cases. Finally, our data confirm the high prevalence of T. rubrum commonly observed in Europe as causal agent of onychomycosis (70,9%, n=1603) followed by T. mentagrophytes complex (20,9%, n=455). T. rubrum and T. mentagrophytes complex are also responsible for the majority of skin infections as they represent respectively 40% (n=386) and 24,75% (n=239) of skin dermatophytosis during the study period. Conclusions The present work has provided recent data on the prevalence of several dermatophytes species circulating in Belgium. Such data is critical for the establishment of therapeutic strategies and measures for prevention and control of dermatophytes infections. Our study confirms the predominance of T. rubrum followed by T. mentagrophytes in the Belgian population but also highlights the emergence of new anthropophilic species such as M. audouinii and T. violaceum as causative agents of tinea capitis in children in relation with African immigration. [less ▲]

Detailed reference viewed: 39 (5 ULg)
Full Text
Peer Reviewed
See detailEpidemiological aspects and genotypic characterization of T.violaceum strains collected during a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULg; Dekkers, Charlotte; GRAIDE, Hélène ULg 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 ▲]

Detailed reference viewed: 28 (5 ULg)
Full Text
Peer Reviewed
See detailOnychomycosis: is it possible to increase the cure rate?
Hayette, Marie-Pierre ULg

in Mycoses (2015, October), 58(suppl 4), 35

Onychomycosis represent about 50% of nails disorders in the world with a very variable prevalence depending of the countries considered. Dermatophytes, non-dermatophyte molds and Candida sp. are the main ... [more ▼]

Onychomycosis represent about 50% of nails disorders in the world with a very variable prevalence depending of the countries considered. Dermatophytes, non-dermatophyte molds and Candida sp. are the main causing agents. A rapid and accurate diagnosis is essential in order to give an adequate treatment to the patient. Generally, a combination of microscopy and culture is used for laboratory diagnosis. However, microscopy does not always allow the distinction between yeasts or filamentous fungi, culture takes generally about a week before identification and the result is compromised if there is contamination by not relevant fungi. Therefore commercially available PCR-based methods have been developed in order to provide a rapid and accurate identification of dermatophytes and yeasts in nails samples. Combination of microscopy and PCR may provide a rapid and specific diagnosis in 2 working days. However this methodology is still not widely used by laboratories because of the high cost. Furthermore, this technology can detect DNA from dead fungi and therefore is not suitable for assessment of treatment efficacy. Onychomycosis therapy depends on different factors such as the causative agent, the number of nails and degree of nail involvement, the type of onychomycosis, potential drug interactions or drug intolerance and a failure to previous treatments. Oral and topical antifungals are mostly used separately or in combination. Oral therapy includes azoles (itraconazole, fluconazole) and/or allylamine (terbinafin), this latter being the most frequently prescribed antifungal for treatment of onychomycosis in North America and Europe. Topical amorolfine and ciclopirox formulations can be used alone in mild cases or in case of intolerance to oral antifungals. However, one of the biggest problems of therapy for onychomycosis is the high frequency of relapse which concerns about 20 to 40% of the patients treated by oral antifungals. Different strategies have been developed to overcome this problem amongst which are: optimization of the dosing regimens (continuous vs pulse therapy) or therapy duration, combination therapy (nail debridement + antifungals, oral + topical drugs, 2 oral drugs), improving drug delivery (use of physical or chemical enhancers, and modification of the pharmacological formulation for increasing drug uptake). Some strategies such as combination therapy (oral + topical) have demonstrated enhanced efficacy and should be recommended in case of poor efficacy of the initial treatment or in case of extended infection. Prophylactic topical therapy implemented after completion of oral treatment has been shown to delay relapse. Preventive measures such as treatment of concurrent tinea pedis and/or infected family members and regular cleaning of bathroom and shower floors can help to reduce the risk of reinfection particularly when a dermatophyte is the causative agent. In conclusion, treatment for onychomycosis is associated with frequent relapse. Consequently, follow-up is mandatory and combination therapy can be necessary in case of relapse or resistance to treatment. Patients should also be aware of the preventive hygiene measures to apply in order to decrease the risk of reinfection. New strategies improving treatment efficacy are promising but their efficacy have still to be demonstrated in comparative clinical trials before their implementation in therapy. [less ▲]

Detailed reference viewed: 22 (0 ULg)
Full Text
Peer Reviewed
See detailDermatophytosis, Trends in Epidemiolgy and Diagnostic Approach
HAYETTE, Marie-Pierre ULg; SACHELI, Rosalie ULg

in Current Fungal Infections report (2015), 9(3), 164-179

Dermatophytes are among the common fungal agents implicated in superficial skin infections. The anthropophilic dermatophyte Trichophyton rubrum is still the most frequent causative agent worldwide but the ... [more ▼]

Dermatophytes are among the common fungal agents implicated in superficial skin infections. The anthropophilic dermatophyte Trichophyton rubrum is still the most frequent causative agent worldwide but the prevalence of several species of dermatophytes varies through different areas around the world. This review summarizes the current status of dermatophytes infection in Europe, Africa, Asia and America and gives an overview of the molecular biology laboratory methods currently available for the diagnosis of dermatomycoses. [less ▲]

Detailed reference viewed: 36 (7 ULg)
Full Text
Peer Reviewed
See detailEpidemiological aspects and genotypic characterization of strains of Microsporum audouinii isolated in the context of a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULg; Dekkers, Charlotte; DARFOUF, Rajae ULg et al

Poster (2015, May)

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

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

Detailed reference viewed: 58 (9 ULg)
Full Text
Peer Reviewed
See detailEpidemiological aspects and genotypic characterization of strains of Microsporum audouinii isolated in the context of a Belgian National survey on anthropophilic tinea
SACHELI, Rosalie ULg; Géron, Bénédicte; Dekkers, Charlotte et al

Poster (2015, April 28)

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

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

Detailed reference viewed: 27 (9 ULg)
Full Text
Peer Reviewed
See detailEpidemiology and clinical reporting of candidaemia in Belgium : a national prospective study (TANSIR trial)
Trouvé, Charlotte; Blot, Stijn; Hayette, Marie-Pierre ULg et al

Poster (2015, April 26)

Objectives The aim of this multicenter study was to gather epidemiological data on candidemia in the Belgian population. Another goal was to determine the time in real life setting for reporting to the ... [more ▼]

Objectives The aim of this multicenter study was to gather epidemiological data on candidemia in the Belgian population. Another goal was to determine the time in real life setting for reporting to the treating physicians of the species involved and its antifungal susceptibility. Methods Prospective study in 29 Belgian hospitals. From March 1st, 2013 till February 28, 2014 the first Candida isolate from each episode of candidemia was included. Identification and susceptibility testing were performed according to local procedures and isolates were sent to the National Reference Lab with a completed case report form. Species identification was checked by MALDI-TOF mass spectrometry (MS) and ITS sequencing in case no reliable identification was obtained by MS. Antifungal susceptibility testing was performed according to EUCAST guidelines. The total number of patient admissions and hospitalization days during the study period was retrieved from each hospital. Results 341 isolates were retrieved from 325 patients (53.2% male, median age 66 years, range 1-94 years) admitted to the ICU (34.4%), medical wards (30.8%), surgical wards (15.2%), onco-haematology (10.6%), pediatrics (3.0%), neonatology (1.7%) and other wards (4.3%). The mean incidence rate of candidemia was 0.42 per 1,000 admissions (range 0.07 to 1.44) and 0.60 per 10,000 patient days (range from 0.11 to 2.03). Candida albicans was the main cause of candidemia (51.9%), followed by Candida glabrata (26.7%), Candida parapsilosis (9.9%), Candida tropicalis (4.4%), Candida guilliermondii (2.6%), Candida dubliniensis (1.5%), Candida lusitaniae (1.2%), Candida krusei (1.2%) and Candida metapsilosis (0.6%). Overall resistance to fluconazole was 6.7% and to anidulafungin 0.6% (2 C. glabrata isolates were echinocandin resistant). Resistance to amphotericin B was detected in 1 C. tropicalis isolate, all C. albicans, C. glabrata and C. parapsilosis isolates remained susceptible to this drug. Resistance to fluconazole ranged from 3.5% in C. albicans, 8.6% in C. glabrata, 5.6% in C. parapsilosis to 35.7% (5/14 isolates) in C. tropicalis. These five C. tropicalis isolates showed cross resistance to voriconazole and posaconazole. MIC values for caspofungin ranged from <0,016 to >8mg/L, with MIC50 of 0.06mg/L and MIC90 of 0.25mg/L. The median time between blood sampling and positivity of the blood culture bottle was 37h07min (Q1-Q3: 25h42min-54h28min). The median time between blood culture positivity and reporting of isolate identification and susceptibility to the treating physician was 29h58min (Q1-Q3: 23h21min-40h34min) and 59h34min (Q1-Q3: 48h28min-75h20min) respectively. Conclusions A large variation in the incidence of candidemia among Belgian hospitals was observed. Resistance to azole drugs remained low but emerging resistance to these drugs among C. tropicalis was noted. Resistance to echinocandins remains rare in Belgian Candida isolates. These data will be further analyzed in order to evaluate the influence of the identification and susceptibility testing method on the time to report results to the treating physicians. [less ▲]

Detailed reference viewed: 33 (3 ULg)
Full Text
Peer Reviewed
See detailEvaluation of a commercially developed semi-automated PCR-surface enhanced raman scattering assay for the detection of Candida species in blood
HAYETTE, Marie-Pierre ULg; WERY, Marie ULg; BOREUX, Raphaël ULg et al

Poster (2015, April 25)

Objectives Microbiological diagnosis of invasive candidiasis is still dependent on culture-based methods. The use of beta-D-glucan antigen detection is included in the EORTC microbiological diagnostic ... [more ▼]

Objectives Microbiological diagnosis of invasive candidiasis is still dependent on culture-based methods. The use of beta-D-glucan antigen detection is included in the EORTC microbiological diagnostic criteria but is rarely available in the clinical labs. On the other hand, PCR-based methods lack standardization. The RenDx Fungiplex® is a new commercially available semi-automated PCR SERS assay designed for the detection of Aspergillus sp. and Candida sp. including the differentiation of resistant strains as C. glabrata, C. krusei and A. terreus. This study was performed for sensitivity and reproducibility testing of the method on 8 different Candida species. Methods The study was conducted on EDTA-blood collected from a healthy donor. Blood samples were spiked with 10 Candida reference strains: C. albicans ATCC 10231, C. albicans NEQAS 1206 and C. albicans NEQAS 2359; C. glabrata ATCC 90030 ; C. krusei ATCC 6258 ; C. tropicalis NEQAS 1036 ; C. guillermondii NEQAS 1035 ; C. parapsilosis ATCC 22019 ; C. lusitaniae NEQAS 1511 and C. dubliniensis IHEM 14280. Spiked samples were diluted at final concentrations ranging from 1 CFU/mL to 1000 CFU/mL. Cultures on Sabouraud dextrose agar were performed in parallel to control yeasts dilutions. DNA extraction was performed by using proteinase K-based method followed by purification on QIAcube automate. The RenDx Fungiplex®kit (Renishaw) was used for the amplification process and the final detection was processed on the SP-1000 sample analyzer. Reproducibility testing was performed on the three C. albicans reference strains by repeating each test 5 times. Results A total of 142 samples were included in the study. A sensitivity of 10 CFU/mL was reached for C. glabrata, C. krusei, C. tropicalis, C. dubliniensis spiked samples while C. lusitaniae and C. tropicalis performed better at 1 CFU/mL. The three tested reference C. albicans strains and C. guillermondii gave the lowest sensitivity (100 CFU/mL). The reproducibility of the assay was 96% Conclusion RenDx Fungiplex®kit allows the detection of the most frequent Candida species responsible for invasive candidiasis in spiked blood samples. The sensitivity of the test is comprised between 10 and 100 CFU/mL for most Candida sp. and reproducibility is very high. This evaluation allows us to consider this commercial kit for inclusion in a clinical study on invasive candidiasis in comparison with non-molecular diagnostic assays. [less ▲]

Detailed reference viewed: 65 (11 ULg)
Full Text
Peer Reviewed
See detailComparison of an In-House Quantitative Real-Time PCR and COBAS ampliprep/Taqman Roche for determination of viral load for HIV type 1 non-b
KAMANGU, Erick; CHATTE, Adawaye; BOREUX, Raphaël ULg et al

in Open access Library Journal (2015), 2(e1402),

Context: The in-house techniques or experimental methods are increasingly recommended for their low-cost reagents for the determination of the Viral Load (VL) in resource-limited settings. The objective ... [more ▼]

Context: The in-house techniques or experimental methods are increasingly recommended for their low-cost reagents for the determination of the Viral Load (VL) in resource-limited settings. The objective of this study was to compare the determination of VL from HIV-1 non-B samples by an in-house technique with the COBAS AmpliPrep/TaqMan version 2.0. Method: In this cross-sectional study, 39 plasma samples from patients infected with HIV type 1 non-B from N’Djamena and Kinshasa were used to determine the VL using the two techniques. Results: The mean values of VL are respectively 4.68 ± 1.26 and 4.58 ± 1.33 log10 RNA copies/ml for the COBAS AmpliPrep/TaqMan assays and the in-house assays. A good correlation (Spearman Correlation) was obtained, with a coefficient (R2) of 0.9452. Conclusion: This study demonstrates that there is no significant difference between the results of VL determined by the COBAS AmpliPrep/TaqMan assays and the in-house assays used. [less ▲]

Detailed reference viewed: 18 (1 ULg)
Full Text
Peer Reviewed
See detailDirect detection of Aspergillus and azole resistance of Aspergillus fumigatus on broncho-alveolar lavage fluid. Validation of a new Aspergillus real-time PCR
Chong, Ga-Lai; Van de Sande, Wendy; Dingemans, Gijs et al

in Journal of Clinical Microbiology (2015)

Introduction Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay consisting of ... [more ▼]

Introduction Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay consisting of two multiplex real-time PCRs: one which identifies the clinically relevant Aspergillus species, and one which detects the TR34, L98H, T289A, Y121F mutations in CYP51A and differentiates susceptible from resistant A. fumigatus strains. Methods The diagnostic performance was tested on 37 bronchoalveolar lavage (BAL) samples from haematology patients and on 40 BAL samples from intensive care unit (ICU) patients using BAL galactomannan ≥1.0 or positive culture as the gold standard for the presence of Aspergillus. Results In the haematology and ICU groups combined, there were 22 BAL samples with IA (2 proven, 9 probable and 11 non-classifiable). Nineteen of the 22 BAL samples were positive according to the gold standard. The optimal cycle threshold value for the presence of Aspergillus was <36. Sixteen of the 19 BAL samples had a positive PCR (2 Aspergillus species and 14 A. fumigatus). This resulted in a sensitivity, specificity, positive and negative predictive value of 88.9%, 89.3%, 72.7% and 96.2% for the haematology group and 80.0%, 93.3%, 80.0% and 93.3% in the ICU group, respectively. The CYP51A real-time PCR confirmed 12 wildtype and 2 resistant strains (1 TR34/L98H and 1 TR46/Y121F/T289A mutant). Conclusion The AsperGenius® multiplex real-time PCR allows for a sensitive and fast detection of Aspergillus species directly in BAL samples. More importantly, this assay detects and differentiates wildtype from resistant strains even if BAL cultures remained negative. [less ▲]

Detailed reference viewed: 22 (3 ULg)
Full Text
Peer Reviewed
See detailGrowth of desferrioxamine deficient Streptomyces mutants through xenosiderophore piracy of airborne fungal contaminations
Arguelles Arias, Anthony ULg; Lambert, Stephany; Martinet, Loïc et al

in FEMS Microbiology Ecology (2015)

Detailed reference viewed: 36 (8 ULg)
Peer Reviewed
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 ▲]

Detailed reference viewed: 78 (18 ULg)
Full Text
Peer Reviewed
See detailDevelopment of a new commercial qPCR assay to detect and differentiate dermatophyte infections of the skin, nails and hair
Dingemans, Gijs; van den Bosch, Mélanie; HAYETTE, Marie-Pierre ULg et al

Poster (2014, May)

Detailed reference viewed: 18 (1 ULg)
Peer Reviewed
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 ▲]

Detailed reference viewed: 80 (22 ULg)