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Robin C.,Henri Mondor Teaching Hospital | Robin C.,University Paris Est Creteil | Alanio A.,Parasitology Mycology Laboratory | Alanio A.,University of Paris Pantheon Sorbonne | And 3 more authors.
Current Opinion in Hematology | Year: 2014

Purpose of review This study focuses on the epidemiology and management of mucormycosis in hematopoietic stem cell transplant patients, a life-threatening mold infection whose incidence has increased over the past decades. Recent findings Mucormycosis may occur in hematopoietic stem cell transplant recipients with severe graft-versus-host disease, steroids, neutropenia, iron overload, diabetes, and malnutrition, or those who received antifungals not active against Mucorales. Its incidence in allogeneic hematopoietic stem cell transplant is around 0.3%. As Mucorales are not susceptible to voriconazole and candins, and as mucormycosis often mimics aspergillosis, it is extremely important to have a precise diagnostic to correctly manage the patient. The reversed halo sign on chest computed tomography has been associated to mucormycosis in neutropenic patients, but is not pathognomonic. Direct fungal identification is crucial. Molecular approaches are developed that may be extremely useful for early diagnosis. Summary Although randomized trials are quite impossible to run, due to the rarity of the disease, the recent numerous data have allowed the elaboration of European guidelines for the management of mucormycosis. Lipid formulations of amphotericin B, and especially liposomal amphotericin B at high doses (5-10mg/kg/day), are the standard treatment, combined with surgery and control of favoring factors. The prognosis is poor, and any delay in the initiation of therapy may impact on outcome. © 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins. Source

Maubon D.,Parasitology Mycology Laboratory | Hamidfar-Roy R.,Medical ICU | Courby S.,Albert Michallon Teaching Hospital | Vesin A.,Joseph Fourier University | And 8 more authors.
Journal of Infection | Year: 2010

Objectives: New molecular methods allow rapid pathogen detection in patients with sepsis, but their impact on treatment decisions remains to be established. We evaluated the therapeutic usefulness of multiplex PCR testing in patients with cancer and sepsis. Methods: 110 patients with cancer and sepsis were included prospectively and underwent LightCycler® SeptiFast (LC-SF) multiplex PCR testing in addition to standard tests. Two independent panels of experts assessed the diagnosis in each patient based on medical record data; only one panel had the LC-SF results. The final diagnosis established by a third panel was the reference standard. Results: The final diagnosis was documented sepsis in 50 patients (55 microorganisms), undocumented sepsis in 54, and non-infectious disease in 6. LC-SF detected 17/32 pathogens recovered from blood cultures (BC) and 11/23 pathogens not recovered from BC; 12 microorganisms were detected neither by BC nor by LC-SF. LC-SF produced false-positive results in 10 cases. The LC-SF results would have significantly improved treatment in 11 (10%) patients and prompted immediate antimicrobial therapy not given initially in 3 patients. Conclusions: In cancer patients with suspected sepsis, LC-SF detected 11/55 (20%) true pathogens not recovered from BCs and would have improved the initial management in 11/110 (10%) patients. © 2010 The British Infection Society. Source

Alanio A.,Parasitology Mycology Laboratory | Alanio A.,University Paris Diderot | Alanio A.,French National Center for Scientific Research | Bretagne S.,Parasitology Mycology Laboratory | And 3 more authors.
Clinical Microbiology and Infection | Year: 2014

PCR assays have not reached the same level of acceptance for the detection of human fungal pathogens as for other micro-organisms, mainly because the low number of micro-organisms challenges the detection limits of PCR. Therefore, whereas meta-analyses focusing on clinical validation suggest interest in adding PCR results to the diagnostic workup for invasive fungal disease (IFD) along with clinical evaluation, CT scans, classical mycology and antigen detection, no consensual PCR method has emerged. Compared with the end-point format of the 1990s, real-time quantitative PCR is a major breakthrough. This format prevents contamination with previously amplified products, provides the yield of amplification, allows for developing consensus procedures and should therefore be the only format used. An internal control is now mandatory to avoid false-negative results. Primer design strongly impacts on the objectives: pan-fungal primers can provide false-positive results due to environmental fungal DNA contamination; conversely, species-specific primers miss infections caused by untargeted fungi. Unresolved issues include the best specimens to be used; serum is currently preferred to blood because of the ease of the DNA extraction step. Work is in progress to establish standards at least for Aspergillus PCR, and the implementation of quality controls should help centres to improve assays. Eventually, the classical analysis of biomarker performance does not consider the evolving risk factors and changing treatments during IFD, which can lead to variable conclusions. New statistical methods such as event history analysis should be considered. © 2014 The Authors. Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases. Source

Even C.,Assistance Publique Hopitaux de Paris AP HP | Bastuji-Garin S.,AP HP | Bastuji-Garin S.,University Paris Est Creteil | Hicheri Y.,Assistance Publique Hopitaux de Paris AP HP | And 11 more authors.
Haematologica | Year: 2011

Patients with acute leukemia who initially survive invasive fungal disease must receive chemotherapy or go on to transplant. Many centers change subsequent chemotherapy to decrease the risk of fungal reactivation. This case-control study compared acute leukemia patients (n=28) who developed a proven or probable fungal disease and survived four weeks later, to patients who did not (n=78), and assessed the impact of fungal disease on the chemotherapy regimens, and overall and event-free survival. Chemotherapy changes (i.e. delays, dose-reduction) were more frequent in the fungal (68%) than in the control group (24%) (P<0.001). Although there was no difference in overall and event-free survival between groups, they were both lower for proven fungal disease cases when compared to controls (HR 2.4, 95% CI 1.1-1.5, and HR 2.9, 95% CI 1.4- 5.6, respectively). Patients with invasive fungal disease, even though they initially survive, undergo significant changes to their chemotherapy therapy. This impacts on the survival of patients with proven fungal disease. ©2011 Ferrata Storti Foundation. Source

Maubon D.,Parasitology Mycology Laboratory | Maubon D.,Joseph Fourier University | Dubosson M.,Parasitology Mycology Laboratory | Chiquet C.,Joseph Fourier University | And 9 more authors.
Investigative Ophthalmology and Visual Science | Year: 2012

PURPOSE. As thenumber of cases of Acanthamoeba spp. keratitis (AK) is constantly growing, new diagnostic tools are needed to confirm and guide ophthalmologists in this clinically problematic diagnosis. Molecular diagnosis is particularly well adapted, although only a few real-time PCR techniques have been described recently. The aim of this study was to develop a new PCR technique for the diagnosis of AK by combining the detection of Acanthamoeba DNA with human DNA, thus allowing an accurate interpretation of the PCR result. METHODS. Different DNA extraction procedures were compared to ensure an optimized amplification of one Acanthamoeba genome. The analytical parameters of this new multiplex Acanthamoeba beta-globin PCR (MAB-PCR) were evaluated. Fourteen eye drops were tested as potential PCR inhibitors. A prospective series of 28 corneal scrapings was subjected to MAB-PCR. RESULTS. The best extraction procedure associated thermalshock pretreatment followed by a manual extraction procedure. The MAB-PCR parameters displayed excellent specificity and sensitivity, with a detection of 0.02 genome of Acanthamoeba. No eye drops were total PCR inhibitors. Of 28 corneal scrapings, 18 were considered true negatives. Seven could not be interpreted because of insufficient scraping material. Three were considered true positives: every patient progressed favorably on specific and reliable treatment. CONCLUSIONs. The MAB-PCR is a new tool to diagnose AK. It allows rapid diagnosis and prompt treatment of this probably underestimated etiology of infectious keratitis. This optimized real-time PCR outperforms the gold standard for Acanthamoeba keratitis diagnosis and it allows a concomitant evaluation of the quality of the corneal scraping, which is necessary for a precise interpretation of the results. © 2012 The Association for Research in Vision and Ophthalmology, Inc. Source

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