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A Multiplex PCR and DNA-Sequencing Workflow on Serum for the Diagnosis and Species Identification for Invasive Aspergillosis and Mucormycosis.
Imbert, S; Portejoie, L; Pfister, E; Tauzin, B; Revers, M; Uthurriague, J; Hernandez-Grande, M; Lafon, M-E; Jubert, C; Issa, N; Dumas, P-Y; Delhaes, L.
  • Imbert S; Centre Hospitalier Universitaire de Bordeaux, Service de Parasitologie-Mycologie, Bordeaux, France.
  • Portejoie L; Université de Bordeaux, Centre de Recherche Cardio-Thoracique de Bordeaux, Pessac, France.
  • Pfister E; Centre Hospitalier Universitaire de Bordeaux, Service de Parasitologie-Mycologie, Bordeaux, France.
  • Tauzin B; Centre Hospitalier Universitaire de Bordeaux, Service de Parasitologie-Mycologie, Bordeaux, France.
  • Revers M; Centre Hospitalier Universitaire de Bordeaux, Plateau Technique de Biologie Moleculaire, Bordeaux, France.
  • Uthurriague J; Centre Hospitalier Universitaire de Bordeaux, Service de Parasitologie-Mycologie, Bordeaux, France.
  • Hernandez-Grande M; Centre Hospitalier Universitaire de Bordeaux, Service de Parasitologie-Mycologie, Bordeaux, France.
  • Lafon ME; Centre Hospitalier Universitaire de Bordeaux, Plateau Technique de Biologie Moleculaire, Bordeaux, France.
  • Jubert C; Centre Hospitalier Universitaire de Bordeaux, Plateau Technique de Biologie Moleculaire, Bordeaux, France.
  • Issa N; Centre Hospitalier Universitaire de Bordeaux, Service d'Hématologie et Cancérologie pédiatrique, Bordeaux, France.
  • Dumas PY; Centre Hospitalier Universitaire de Bordeaux, Service de Médecine Intensive - Réanimation, Bordeaux, France.
  • Delhaes L; Centre Hospitalier Universitaire de Bordeaux, Service d'Hématologie et Thérapie cellulaire, Pessac, France.
J Clin Microbiol ; 61(1): e0140922, 2023 01 26.
Article in English | MEDLINE | ID: covidwho-2193443
ABSTRACT
There has been significant increase in the use of molecular tools for the diagnosis of invasive aspergillosis (IA) and mucormycosis. However, their range of detection may be too limited as species diversity and coinfections are increasing. Here, we aimed to evaluate a molecular workflow based on a new multiplex PCR assay detecting the whole Aspergillus genus and the Mucorales order followed by a species-specific PCR or a DNA-sequencing approach for IA and/or mucormycosis diagnosis and species identification on serum. Performances of the MycoGENIE Aspergillus spp./Mucorales spp. duplex PCR kit were analyzed on a broad range of fungal strains and on sera from high-risk patients prospectively over a 12-month period. The kit allowed the detection of nine Aspergillus species and 10 Mucorales (eight genera) strains assessed. No cross-reactions between the two targets were observed. Sera from 744 patients were prospectively analyzed, including 35 IA, 16 mucormycosis, and four coinfections. Sensitivity varies from 85.7% (18/21) in probable/proven IA to 28.6% (4/14) in COVID-19-associated pulmonary aspergillosis. PCR-positive samples corresponded to 21 A. fumigatus, one A. flavus, and one A. nidulans infections. All the disseminated mucormycosis were positive in serum (14/14), including the four Aspergillus coinfections, but sensitivity fell to 33.3% (2/6) in localized forms. DNA sequencing allowed Mucorales identification in serum in 15 patients. Remarkably, the most frequent species identified was Rhizomucor pusillus (eight cases), whereas it is barely found in fungal culture. This molecular workflow is a promising approach to improve IA and mucormycosis diagnosis and epidemiology.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Aspergillosis / Coinfection / Invasive Fungal Infections / COVID-19 / Mucorales / Mucormycosis Type of study: Diagnostic study / Experimental Studies / Prognostic study / Randomized controlled trials Limits: Humans Language: English Journal: J Clin Microbiol Year: 2023 Document Type: Article Affiliation country: Jcm.01409-22

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Aspergillosis / Coinfection / Invasive Fungal Infections / COVID-19 / Mucorales / Mucormycosis Type of study: Diagnostic study / Experimental Studies / Prognostic study / Randomized controlled trials Limits: Humans Language: English Journal: J Clin Microbiol Year: 2023 Document Type: Article Affiliation country: Jcm.01409-22