Laurence Millon1,2, Denis Caillot3, Ana Berceanu4, Stéphane Bretagne5,6,7, Fanny Lanternier5,7,8, Florent Morio9,10, Valérie Letscher-Bru11, Frédéric Dalle12,13, Blandine Denis14, Alexandre Alanio5,6,7, David Boutoille15, Marie Elisabeth Bougnoux16,17, Françoise Botterel18,19, Taieb Chouaki20,21, Amandine Charbonnier22, Florence Ader23, Damien Dupont24, Anne Pauline Bellanger1,2, Steffi Rocchi1,2, Emeline Scherer1,2, Houssein Gbaguidi-Haore2,25, Raoul Herbrecht26,27. 1. Laboratoire de Parasitologie-Mycologie, Centre hospitalier universtaire (CHU) Besançon, Besançon, France. 2. Unité mixte de recherche (UMR) 6249 Centre national de la recherche scientifique (CNRS) Chrono-Environnement, Univ Bourgogne Franche-Comté, Besançon, France. 3. Department of Clinical Hematology, CHU Dijon, Dijon, France. 4. Service d'Hematologie, CHU Besançon, Besançon, France. 5. Institut Pasteur, CNRS, Molecular Mycology Unit, National Reference Center for Invasive Mycoses and Antifungals, UMR 2000, Paris, France. 6. Laboratoire de Parasitologie-Mycologie, Hôpital Saint Louis, AP-HP, Paris, France. 7. Université de Paris, Paris, France. 8. Necker Pasteur Center for Infectious Diseases and Tropical Medicine, Hôpital Necker Enfants malades, AP-HP, Instituts Hôpitaux Universitaires (IHU) Imagine, Paris, France. 9. Laboratoire de Parasitologie-Mycologie, CHU Nantes, Nantes, France. 10. Département de Parasitologie et Mycologie Médicale, EA1155 - IICiMed, Nantes Université, Nantes, France. 11. Laboratoire de Parasitologie et de Mycologie Médicale, Hôpitaux Universitaires de Strasbourg, France. 12. Laboratoire de Parasitologie-Mycologie, Plateforme de Biologie Hospitalo-Universitaire Gérard Mack, Dijon, France. 13. UMR PAM Univ Bourgogne Franche-Comté, AgroSup Dijon, Equipe Vin, Aliment, Microbiologie, Stress, Dijon, France. 14. Infectious Diseases Department, APHP, Saint-Louis Hospital, Paris, France. 15. Unité Maladies Infectieuses et Tropicales, CHU Nantes, Nantes, France. 16. Parasitology-Mycology Unit, Necker Enfants Malades Hospital, APHP, Paris, France. 17. Fungal Biology and Pathogenicity Unit, INRA USC 2019, Institut Pasteur, Paris, France. 18. EA Dynamyc 7380 UPEC, ENVA, Faculté de Médecine, Créteil, France. 19. Unité de Parasitologie, Mycologie, Département de Virologie, Bactériologie-Hygiène, Mycologie-Parasitologie, DHU VIC, CHU Henri Mondor, Créteil, France. 20. Laboratoire de Parasitologie et Mycologie Médicales, Centre de Biologie Humaine, CHU Amiens Picardie, Amiens, France. 21. Equipe AGIR: Agents Infectieux, Résistance et Chimiothérapie UR4294, Université de Picardie Jules Verne, Amiens, France. 22. Department of Clinical Hematology and Cellular Therapy, Amiens University Medical Center, Amiens, France. 23. Hospices Civils de Lyon, Département des Maladies Infectieuses et Tropicales, F-69004, Lyon, France. 24. Institut des Agents Infectieux, Parasitologie Mycologie, Hôpital de la Croix-Rousse, Hospices Civils de Lyon, Lyon, France. 25. Infection Control Department, CHU Besançon, Besançon, France. 26. Université de Strasbourg, INSERM, IRFAC UMR-S1113, Strasbourg, Franceand. 27. Service d'Hématologie, Institut de Cancérologie Strasbourg Europe (ICANS), Strasbourg, France.
Abstract
BACKGROUND: Early diagnosis and prompt initiation of specific antifungal treatment are essential for improving the prognosis of mucormycosis. We aimed to assess the performance of serum Mucorales quantitative polymerase chain reaction (qPCR) for the early diagnosis and follow-up of mucormycosis. METHODS: We prospectively enrolled 232 patients with suspicion of invasive mold disease, evaluated using standard imaging and mycological procedures. Thirteen additional patients with proven or probable mucormycosis were included to analyze DNA load kinetics. Serum samples were collected twice-a-week for Mucorales qPCR tests targeting the Mucorales genera Lichtheimia, Rhizomucor, and Mucor/Rhizopus. RESULTS: The sensitivity was 85.2%, specificity 89.8%, and positive and negative likelihood ratios 8.3 and 0.17, respectively in this prospective study. The first Mucorales qPCR-positive serum was observed a median of 4 days (interquartile range [IQR], 0-9) before sampling of the first mycological or histological positive specimen and a median of one day (IQR, -2 to 6) before the first imaging was performed. Negativity of Mucorales qPCR within seven days after liposomal-amphotericin B initiation was associated with an 85% lower 30-day mortality rate (adjusted hazard ratio = 0·15, 95% confidence interval [.03-.73], P = .02). CONCLUSIONS: Our study argues for the inclusion of qPCR for the detection of circulating Mucorales DNA for mucormycosis diagnosis and follow-up after treatment initiation. Positive results should be added to the criteria for the consensual definitions from the European Organization for the Research and Treatment of Cancer/Mycoses Study Group Education and Research Consortium (EORTC/MSGERC), as already done for Aspergillus PCR.
BACKGROUND: Early diagnosis and prompt initiation of specific antifungal treatment are essential for improving the prognosis of mucormycosis. We aimed to assess the performance of serum Mucorales quantitative polymerase chain reaction (qPCR) for the early diagnosis and follow-up of mucormycosis. METHODS: We prospectively enrolled 232 patients with suspicion of invasive mold disease, evaluated using standard imaging and mycological procedures. Thirteen additional patients with proven or probable mucormycosis were included to analyze DNA load kinetics. Serum samples were collected twice-a-week for Mucorales qPCR tests targeting the Mucorales genera Lichtheimia, Rhizomucor, and Mucor/Rhizopus. RESULTS: The sensitivity was 85.2%, specificity 89.8%, and positive and negative likelihood ratios 8.3 and 0.17, respectively in this prospective study. The first Mucorales qPCR-positive serum was observed a median of 4 days (interquartile range [IQR], 0-9) before sampling of the first mycological or histological positive specimen and a median of one day (IQR, -2 to 6) before the first imaging was performed. Negativity of Mucorales qPCR within seven days after liposomal-amphotericin B initiation was associated with an 85% lower 30-day mortality rate (adjusted hazard ratio = 0·15, 95% confidence interval [.03-.73], P = .02). CONCLUSIONS: Our study argues for the inclusion of qPCR for the detection of circulating Mucorales DNA for mucormycosis diagnosis and follow-up after treatment initiation. Positive results should be added to the criteria for the consensual definitions from the European Organization for the Research and Treatment of Cancer/Mycoses Study Group Education and Research Consortium (EORTC/MSGERC), as already done for Aspergillus PCR.