Comparison 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; BOREUX, Raphaël ; LEVAUX, Laetitia et alPoster (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 ▲] Detailed reference viewed: 109 (5 ULg) FATAL ALVEOLAR ECHINOCOCCOSIS OF THE LUMBAR SPINEKEUTGENS, Aurore ; SIMONI, Paolo ; DETREMBLEUR, Nancy et alin Journal of Clinical Microbiology (2012) 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 ▲] Detailed reference viewed: 34 (11 ULg) Cerebral aspergillosis in immunocompromised patient successfully treated with voriconazoleHUWART, Aline ; ; HENROTEAUX, Adrienne et alPoster (2012, November 08) Cerebral aspergillosis is a severe invasive mycosis occurring in immunocompromised patients. This pathology is associated with a high rate of mortality and is a current complication of pulmonary invasive ... [more ▼] Cerebral aspergillosis is a severe invasive mycosis occurring in immunocompromised patients. This pathology is associated with a high rate of mortality and is a current complication of pulmonary invasive aspergillosis. We report the case of a 44-year-old immunocompromised male with a recent history of oropharyngeal carcinoma. At his admission the patient presented with fever and confusion. Imaging revealed the presence of a cerebral abscess combined with lung infiltrates. During hospitalization and despite a broad-spectrum antibiotic regimen his condition worsened. A thin needle aspiration of the abscess was performed for diagnosis purpose. Histological examination of the tissue showed septate and branched hyphae with 45° angles suggestive of Aspergillus. A real-time PCR specific for the detection of Aspergillus sp. was carried out and confirmed the fungal etiology of the abscess. Rare colonies of A. fumigatus were isolated a few days later. The diagnosis of invasive pulmonary aspergillosis complicated by a cerebral dissemination was confirmed. Antifungal treatment based on voriconazole 4 mg/kg q12h was introduced and the dosage was successfully increased up to 5 mg/kg q12h by drug monitoring. This case highlights the usefulness of the Aspergillus PCR for the rapid identification of hyphae in tissue biopsies (or in the event of negative culture), and the importance of therapeutic drug monitoring in treatment by voriconazole. [less ▲] Detailed reference viewed: 19 (7 ULg) Direct 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 ; ; DESCY, Julie et alin 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 ▲] Detailed reference viewed: 22 (5 ULg) Acute cholecystitis with Listeria monocytogenesDESCY, Julie ; De Mol, Patrick ; HAYETTE, Marie-Pierre et alin 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 ▲] Detailed reference viewed: 10 (2 ULg) Echinoccose alvéolaire : l'expérience liégeoiseHayette, Marie-Pierre ![]() Conference (2012, June 09) Detailed reference viewed: 17 (0 ULg) Validation of new automated chemiluminescent assay for serodiagnosis of human parvovirus B19 infectionHuynen, Pascale ; Toussaint, Françoise ; Melin, Pierrette et alPoster (2010, December) Detailed reference viewed: 30 (5 ULg) Filamentous fungi recovered from the water distribution system of a Belgian university hospitalHayette, Marie-Pierre ; Christiaens, Geneviève ; Mutsers, Jacques et alin Medical Mycology (2010), 48(7), 969-974 A study was carried out over a 4-month winter period in order to assess the presence of filamentous fungi in the water distribution system of the University Hospital of Liège. A total of 197 hot and cold ... [more ▼] A study was carried out over a 4-month winter period in order to assess the presence of filamentous fungi in the water distribution system of the University Hospital of Liège. A total of 197 hot and cold water samples were collected from the main water supply lines and from the taps at three different hospital sites. Overall, filamentous fungi were recovered from 55% and 50% of the main water distribution system and tap water samples, respectively, with a mean of 3.5 ± 1.5 colony forming units per 500 ml water. Nine different genera were identified, all belonging to the Hyphomycetes class. Aspergillus spp. were recovered from 6% of the samples of the water distribution system and A. fumigatus was the most frequently recovered species (66.6%). However, this species was not isolated from water taps. Fusarium spp. was predominant at one site, where it was found in 28% of tap water samples. No Aspergillus spp. but some Fusarium spp. isolates were identified in samples collected from high-risk units. Filters were introduced at the point-of-use in the haematology unit after completion of the study. The findings of the present study confirm the need for further documented studies to evaluate the safety of the hospital water system and to define new preventive measures. [less ▲] Detailed reference viewed: 47 (7 ULg) Aspergillus: still the same as before?Hayette, Marie-Pierre ![]() Conference (2010, October 07) La présentation fait le point sur les évolutions touchant les Aspergillus depuis une vingtaine d'années sur les plan taxonomique, épidémiologique, clinique et thérapeutique. Detailed reference viewed: 15 (3 ULg) Comparative studies for the serodiagnosis of Chlamydophila and Mycoplasma pneumoniae infectionsHuynen, Pascale ; Goebel, Marie-Rose ; Meex, Cécile et alin ESCMID (Ed.) Abstract book of the 20th ECCMID (2010, April) Detailed reference viewed: 29 (7 ULg) Travaux pratiques de microbiologie généraleHAYETTE, Marie-Pierre ; HUYNEN, Pascale ; MEEX, Cécile ![]() Learning material (2010) Le syllabus reprend les méthodologies utilisées pour l'étude des bactéries, levures, virus. Il décrit les réactions sérologiques et les principes de biologie moléculaire appliqués au diagnostic ... [more ▼] Le syllabus reprend les méthodologies utilisées pour l'étude des bactéries, levures, virus. Il décrit les réactions sérologiques et les principes de biologie moléculaire appliqués au diagnostic microbiologique. [less ▲] Detailed reference viewed: 707 (26 ULg) Spectrométrie de masse MALDI-TOF en bactériologie clinique ou comment identifier une bactérie en une minuteDescy, Julie ; Meex, Cécile ; Melin, Pierrette et alin Revue Médicale de Liège (2010), 65(Suppl. Synthèse 2010), 29-34 The major application of MALDI-TOTOF mass spectrometry in clinical microbiology is the bacterial identification based on the analysis of all their proteins (ribosomal and membrane-associated proteins ... [more ▼] The major application of MALDI-TOTOF mass spectrometry in clinical microbiology is the bacterial identification based on the analysis of all their proteins (ribosomal and membrane-associated proteins). This technology allows the identification of most of bacteria within a few minutes. The method is fast, accurate, reliable and cost-effective by comparison to conventional phenotypic techniques. Other applications of MALDI-TOF mass spectrometry are still under development, as the detection of bacterial toxins or resistance mechanisms to antimicrobial agents. [less ▲] Detailed reference viewed: 513 (13 ULg) Mucormycose invasive du poumon et du rachis dorsal.De Pasqual, Aurelie ; Deprez, Manuel ; Ghaye, Benoît et alin Revue Médicale de Liège (2008), 63 Nous rapportons le cas d'un patient de 67 ans atteint d'un syndrome myélodysplasique et qui a développé une mucormycose pulmonaire avec extension tout à fait exceptionnelle vers le rachis dorsal ... [more ▼] Nous rapportons le cas d'un patient de 67 ans atteint d'un syndrome myélodysplasique et qui a développé une mucormycose pulmonaire avec extension tout à fait exceptionnelle vers le rachis dorsal responsable d'un paraplégie aiguë. Après échec d'un traitement probabiliste anti-aspergillaire, c'est finalement l'analyse des prélèvements obtenus lors de la laminectomie décompressive qui a fourni le diagnostic mycologique. En raison d'une altération majeure de l'état général, la lobectomie prévue n'a pu être réalisée et malgré l'adaptation du traitement antifongique (Abelcet, Posaconazole), le patient est décédé. La mucormycose (ou zygomycose) pulmonaire est une infection fongique peu commune qui touche essentiellement les patients immuno-déprimés. Le champignon pathogène fait partie des zygomycètes dont la caractéristique principale est la capacité d'angio-invasion. L'invasion périneurale est une autre voie de propagation récemment mise en évidence. Les difficultés thérapeutiques associées à cette pathologie sont liées au terrain d'immunodépression, aux difficultés d'obtenir rapidement un diagnostic précis ainsi qu'à l'absence de sensibilité du Mucor aux antifongiques récemment introduits (V-Fend, Cancidas). Ceci souligne le risque inhérent à un traitement antifongique empirique par ces agents et la nécessité d'un prélèvement biopsique précoce en cas de non-réponse au traitement. [less ▲] Detailed reference viewed: 177 (11 ULg) A Patient with Hiv Infection, Cough, Asthenia, and Fever; ; et al in Clinical Infectious Diseases : An Official Publication of the Infectious Diseases Society of America (2007), 45(5), 662-3559-600 Detailed reference viewed: 14 (7 ULg) Diagnostic moléculaire de la malariaHayette, Marie-Pierre ![]() Scientific conference (2007, May 30) Le cours porte sur les intérêts et limites des techniques de biologie moléculaire appliquées au diagnostic du paludisme. Detailed reference viewed: 15 (3 ULg) Evaluation of a new commercial real time PCR for the detection of Aspergillus spp. in serum and respiratory samplesHayette, Marie-Pierre ; Meex, Cécile ; Boreux, Raphaël et alPoster (2007, April) Objectives. Diagnosis of invasive aspergillosis is still disappointing and often delayed because of the lack of sensitivity of diagnostic tools. DNA detection based-methods have been developed, but differ ... [more ▼] Objectives. Diagnosis of invasive aspergillosis is still disappointing and often delayed because of the lack of sensitivity of diagnostic tools. DNA detection based-methods have been developed, but differ widely and comparisons are difficult to assess. The objective of the study is to compare a new commercial real-time PCR kit, affigene® Aspergillus tracer assay, with an in house nested PCR targeting 18S rRNA Aspergillus sp. gene. Methods. Twelve patients at risk for invasive aspergillosis were included in the study. They were classified to have possible (5 cases), probable (1 case) or proven (6 cases) invasive aspergillosis following E.O.R.T.C. criteria. Fifteen serum and respiratory paired samples were collected. The DNA extraction was performed by using the QIAmp DNA mini kit® (Qiagen, Germany). All samples were tested by both PCR assays and respiratory samples were cultured. Results. Respiratory samples. A. fumigatus, A. niger and A. flavus were isolated from 10/15 samples; both PCR methods were positive for these samples except one that was positive for affigene® and equivocal for the nested PCR. The real-time PCR assay reported cycle thresholds ranging from 25 to 38. Three of the five culture-negative samples were negative by both PCR methods; one of three was negative in affigene® assay and equivocal by nested PCR; the last sample was positive in affigene® assay and negative by nested PCR. Serum. Thirteen of fifteen blood samples were negative by both PCR methods. One sample was equivocal by nested PCR and was inhibited in affigene® assay despite a culture-positive paired respiratory sample. The last case was inhibited by the real-time PCR assay and negative by nested PCR. Nor the nested PCR, nor affigene® assay could detect any Aspergillus DNA in serum. In total, there was 93% of agreement between the two PCR assays. Conclusion. Both methods are in good agreement and can detect at least three different species of Aspergillus. However, the sensitivity of both assays does not permit the detection of Aspergillus DNA in serum. affigene® assay can easy replace the “in house” assay: it allows a fast and standardized detection of Aspergillus sp. DNA in respiratory samples without inconvenient due to the handling of PCR products. [less ▲] Detailed reference viewed: 47 (4 ULg) Epidémie étrange à Absidia corymbifera en centre de brûlésJACQUEMIN, Denise ; CHRISTIAENS, Geneviève ; HAYETTE, Marie-Pierre et alConference (2007) Detailed reference viewed: 15 (1 ULg) Disseminated Penicillium marneffei infection contrated in ChinaHayette, Marie-Pierre ; ; Meex, Cécile et alPoster (2007) Penicillium marneffei infection is a rare fungal disease that cause significant disease in immunosuppressed patients. The geographical distribution of this dimorphic fungus is restricted to Asia ... [more ▼] Penicillium marneffei infection is a rare fungal disease that cause significant disease in immunosuppressed patients. The geographical distribution of this dimorphic fungus is restricted to Asia, Southeast and Far East, where the disease is considered as an indicator of acquired immunodeficiency syndrome (AIDS). Case report. A 42-year old Congolese woman living in Lubumbashi was admitted at the university hospital of Liège for exploration of a general status impairment. She experienced for three months spiking fever, weight loss, productive cough with bloody expectorations and progressive dyspnoea. She reported also to have non-bloody mild diarrhoea with abdominal pain. The HIV antibody status was positive with a low CD4 T lymphocytes count (28/µl). Pulmonary infiltrates were visualized on chest radiography and the computed tomography revealed the presence of a severe pneumopathy characterised by bilateral micronodular lesions. Mediastinal polyadenopathies associated with hepato- and splenomegaly were also highlighted. Bronchoscopy was performed and bronchial aspirations revealed the presence of numerous leucocytes with the presence of intracellular Gram positive organisms suggestive of yeasts. Ziehl, Giemsa and Gomori-Grocott staining were also performed. Ziehl staining was negative. The morphological aspect given by Giemsa staining excluded infection and the PCR specific for T. gondii B1 gene was negative. However, Gomori-Grocott staining revealed the presence of intracellular oval, elongated, sausage-shaped cells with a single transverse septum (3 to 5 µm). Penicillium marneffei was isolated from blood culture and respiratory samples. Intraveinous amphotericin B treatment was administrated during 15 days followed by itraconazole oral administration (200 mg/j). The antimycotic treatment improved the patient condition and despite other clinical troubles she was prematurely discharged because of financial problems. Conclusion. Opportunistic agents involved in HIV-infected patients differ in Africa and Asia and it is important to be able to make a rapid diagnosis with the aid of an experienced laboratory. [less ▲] Detailed reference viewed: 40 (1 ULg) Pericardite aspergillaire avec tamponnade cardiaque et syndrome hemophagocytaire: un cas non classique d'immunodeficienceDelcroix, Geoffrey ; ; Hustinx, Roland et alin Revue Médicale de Liège (2006), 61(10), 713-8 We report the case of a 31-year-old woman who died in a context of haemophagocytic syndrome with multiple opportunist infections:viral, mycobacterial and fungal. To our knowledge, this is the tenth case ... [more ▼] We report the case of a 31-year-old woman who died in a context of haemophagocytic syndrome with multiple opportunist infections:viral, mycobacterial and fungal. To our knowledge, this is the tenth case of invasive aspergillosis manifested by an aspergillus pericarditis with cardiac tamponade. Search for HIV infection, neoplasia, haematological malignancies was negative. In addition, the patient carried on a nonviral, non-ethylic cirrhosis, the etiology of which remained unknow. We will particularly develop the aspergillus pericarditis with cardiac tamponade and the haemophagocytic syndrome. [less ▲] Detailed reference viewed: 114 (11 ULg) Molecular epidemiology of extended-spectrum beta-lactamase-producing Enterobacteriaceae colonizing the digestive tract of patients admitted to intensive care units in a Belgian university hospitalCHRISTIAENS, Geneviève ; Damas, Pierre ; et alin American Society of Microbiology (Ed.) Program and Abstracts of the 46th Intersciences Conference on Antimicrobial Agents and Chemotherapy (2006, September) Detailed reference viewed: 18 (3 ULg) |
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