title: Early diagnosis and monitoring of mucormycosis by detection of circulating DNA in serum: retrospective analysis of 44 cases collected through the French Surveillance Network of Invasive Fungal Infections (RESSIF).
abstract: The main objective of this study was to assess the diagnostic performance of a set of three Mucorales quantitative PCR assays in a retrospective multicentre study. Mucormycosis cases were recorded thanks to the French prospective surveillance programme (RESSIF network). The day of sampling of the first histological or mycological positive specimen was defined as day 0 (D0). Detection of circulating DNA was performed on frozen serum samples collected from D-30 to D30, using quantitative PCR assays targeting Rhizomucor, Lichtheimia, Mucor/Rhizopus. Forty-four patients diagnosed with probable (n = 19) or proven (n = 25) mucormycosis were included. Thirty-six of the 44 patients (81%) had at least one PCR-positive serum. The first PCR-positive sample was observed 9 days (range 0-28 days) before diagnosis was made using mycological criteria and at least 2 days (range 0-24 days) before imaging. The identifications provided with the quantitative PCR assays were all concordant with culture and/or PCR-based identification of the causal species. Survival rate at D84 was significantly higher for patients with an initially positive PCR that became negative after treatment initiation than for patients whose PCR remained positive (48% and 4%, respectively; p <10-6). The median time for complete negativity of PCR was 7 days (range 3-19 days) after initiation of l-AmB treatment. Despite some limitations due to the retrospective design of the study, we showed that Mucorales quantitative PCR could not only confirm the mucormycosis diagnosis when other mycological arguments were present but could also anticipate this diagnosis. Quantification of DNA loads may also be a useful adjunct to treatment monitoring.
authors.affiliation: Laboratoire de Parasitologie Mycologie, Centre Hospitalier Régional Universitaire, Besançon, France; Laboratoire Chrono-environnement UMR6249, Université Bourgogne Franche-Comté/CNRS, Besançon, France. Electronic address: lmillon@chu-besancon.fr.
authors.forename: L
authors.initials: L
authors.lastname: Millon
authors.affiliation: Département d'Oncologie et Hématologie, Hôpitaux Universitaires de Strasbourg, France.
authors.forename: R
authors.initials: R
authors.lastname: Herbrecht
authors.affiliation: Laboratoire de Parasitologie Mycologie, Centre Hospitalier Régional Universitaire, Besançon, France; Laboratoire Chrono-environnement UMR6249, Université Bourgogne Franche-Comté/CNRS, Besançon, France.
authors.forename: F
authors.initials: F
authors.lastname: Grenouillet
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Nantes, France; Département de Parasitologie et Mycologie Médicale, Université de Nantes, Nantes Atlantique Universités, EA1155-IICiMed, Faculté de Pharmacie, Nantes, France.
authors.forename: F
authors.initials: F
authors.lastname: Morio
authors.affiliation: Centre Hospitalier Universitaire APHP-Saint Louis Paris, France; Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Institut Pasteur, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, France.
authors.forename: A
authors.initials: A
authors.lastname: Alanio
authors.affiliation: Laboratoire de Mycologie Médicale, Plateau Technique de Microbiologie, Hôpitaux Universitaires de Strasbourg, France; Institut de Parasitologie et de Pathologie Tropicale, EA7292, Fédération de Médecine Translationnelle, Université de Strasbourg, France.
authors.forename: V
authors.initials: V
authors.lastname: Letscher-Bru
authors.affiliation: Laboratoire de Parasitologie-Mycologie Centre Hospitalier Universitaire Toulouse, France.
authors.forename: S
authors.initials: S
authors.lastname: Cassaing
authors.affiliation: Centre Hospitalier Universitaire Amiens, France; EA 4666-Centre Universitaire de Recherche en Santé CURS, CAP-Santé (FED 4231), Université de Picardie Jules Verne, France.
authors.forename: T
authors.initials: T
authors.lastname: Chouaki
authors.affiliation: Centre Hospitalier Universitaire Poitiers, France.
authors.forename: C
authors.initials: C
authors.lastname: Kauffmann-Lacroix
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Gabriel Montpied, Clermont-Ferrand, France.
authors.forename: P
authors.initials: P
authors.lastname: Poirier
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Reims, France; Unité MEDyC, CNRS UMR7369, Université Reims Champagne-Ardenne, Reims, France.
authors.forename: D
authors.initials: D
authors.lastname: Toubas
authors.affiliation: Centre Hospitalier Régional Orléans, France.
authors.forename: O
authors.initials: O
authors.lastname: Augereau
authors.affiliation: Laboratoire Chrono-environnement UMR6249, Université Bourgogne Franche-Comté/CNRS, Besançon, France.
authors.forename: S
authors.initials: S
authors.lastname: Rocchi
authors.affiliation: Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Institut Pasteur, Paris, France.
authors.forename: D
authors.initials: D
authors.lastname: Garcia-Hermoso
authors.affiliation: Centre Hospitalier Universitaire APHP-Saint Louis Paris, France; Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Institut Pasteur, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Bretagne
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015897:Comorbidity; D004271:DNA, Fungal / Q000097:blood; D005260:Female; D005602:France / Q000453:epidemiology; D016469:Fungemia; D006801:Humans; D008297:Male; D008875:Middle Aged; D009090:Mucorales / Q000235:genetics*; D009091:Mucormycosis / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology* / Q000628:therapy; D011159:Population Surveillance; D012189:Retrospective Studies; D016019:Survival Analysis
chemical_list: D004271:DNA, Fungal
keywords: Diagnosis; Mucorales; monitoring; mucormycosis; quantitative PCR