Evaluation of a new Histoplasma spp. reverse transcriptase quantitative PCR assay

Autor: Alanio, Alexandre, Gits-Muselli, Maud, Lanternier, Fanny, Sturny-Leclère, Aude, Benazra, Marion, Hamane, Samia, Rodrigues, Anderson Messias, Garcia-Hermoso, Dea, Lortholary, Olivier, Dromer, Françoise, Bretagne, Stéphane, French Mycoses Study Group, The
Přispěvatelé: Mycologie moléculaire - Molecular Mycology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Cité (UPCité), Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals (CNRMA), Institut Pasteur [Paris] (IP), Centre d'infectiologie Necker-Pasteur [CHU Necker], Institut Pasteur [Paris] (IP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Federal University of Sao Paulo (Unifesp), The authors report no funding for this study., The French mycoses study group is composed of: Marine Gosset Woimant, Geneviève blanchard (Centre Hospitaler de Pontoise, Pontoise), Souad Silhadi (Centre Hospitalier de Chambéry, Chambéry), Nicolas Vignier, Aurelia Pitsch, Kaoutar Jidar (Centre Hospitalier de Melun, Melun), Nicolas Traversier (Centre Hospitalier Felix Guyon, La Réunion), Didier Poisson, Claire Lecointre (Centre Hospitalier Régional d'Orléans, Orléans), Françoise Foulet, Françoise Botterel, Nawel Ait Ammar, Amsellem (CHU Henri Mondor, Créteil), Frederic Gabriel (CHU de Bordeau-GH Pellegrin, Bordeaux), Philipe Poirier (CHU de Clermont Ferrand, Clermont-Ferrand), Marjorie Cornu, Severine Loridant (CHU de Lille, Lille), Florent Morio, David Boutoille, Fakhri Jeddi (CHU de Nantes, Nantes), Lilia Hasseine (CHU de Nice, Nice), Rachida Ouissa (CHU de Pointe à Pitre, Guadeloupe), Dominique Toubas (CHU de Reims, Reims), Eric Bailly, Guillaume Désoubeaux (CHU deTours), Emily Ronez (Hôpital Ambroise Paré, Boulogne), Guillaume Foulon, Sebastien Lefrançois (Hôpital Américain, Neuilly), Christine Bonnal (Hôpital Bichat, Paris), André Paugam, Maxime Dougados (Hôpital Cochin, Paris), Marine Desroches (Hôpital d'Instruction des Armées Percy, Clamart), Hélène Barazzutti, Nicolas Paleiron (Hôpital d'Instruction des Armées, Toulon), Meja rabodonirina (Hôpital de la Croix Rousse, Lyon), Emilie Catherinot, Emilie Cardot-Martin, Chrisian Hiesse, Hélène Salvator (Hôpital Foch, Suresnes), Claire Aguilar, Anne Gigandon, Thomas de Montpreville (Hôpital Marie Lannelongue, Le Plessis-Robinson), Marie-Elisabeth Bougnoux, Emilie Sitterlé (Hôpital Necker Enfants Malades, Paris), Arnaud Fekkar, Sébastien Imbert, Alexandre Bleibtreu (Hôpital Pité Salpétrière), Yaye Senghor (Hôpital Saint Joseph, Paris), Blandine Denis, Jean-Michel Molina, Geoffroy Liegeon, Anne-Lise Munnier, Marion Malphettes (Hôpital Saint Louis, Paris), Julie Denis (Hôpitaux Civils de Strasbourg, Strasbourg), Alain Berlioz-Arthaud (Institut Pasteur de Bangui, Bangui, République centrafricaine), Franciska Lange (Centre Hospitalier Sallanches-Chamonix, Sallanches), Myriam Chiaruzzi, Loic Epelboin (Centre Hospitalier Andrée Rosemon, Cayenne, Guyane Française), The authors are grateful to the technical staff of the mycology-parasitology laboratory at Hospital Saint Louis, Paris, France for handling specimens and performing this assay routinely, in particular, Elodie Da Silva, Thierry Pautet, Dieyenaba Siby-Diakite, Sarah seng and Julie Bui, who performed optimization experiments., Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université de Paris (UP), Institut Pasteur [Paris], Institut Pasteur [Paris]-CHU Necker - Enfants Malades [AP-HP]
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Journal of Molecular Diagnostics
Journal of Molecular Diagnostics, 2021, 23 (6), pp.698-709. ⟨10.1016/j.jmoldx.2021.02.007⟩
Journal of Molecular Diagnostics, American Society for Investigative Pathology (ASIP), In press, ⟨10.1016/j.jmoldx.2021.02.007⟩
ISSN: 1525-1578
DOI: 10.1016/j.jmoldx.2021.02.007⟩
Popis: International audience; Laboratory diagnosis of histoplasmosis is based on various methods including microscopy, culture, antigen and DNA detection of Histoplasma capsulatum var. capsulatum (Hcc) or H. capsulatum var. duboisii (Hcd). To improve sensitivity of existing quantitative PCR assays, we developed a new reverse transcriptase qPCR (RTqPCR) assay allowing amplification of whole nucleic acids of Histoplasma spp.. and validated on suspected cases.The limit of detection was 20 copies and the specificity against 114 fungal isolates/species was restricted to Histoplasma spp.. Whole nucleic acids of 1,319 prospectively collected consecutive samples from 907 patients suspected of histoplasmosis were tested routinely between May 2015 and May 2019 in parallel with standard diagnostic procedures performed in parallel. 44 had proven histoplasmosis due to Hcc (n=40) or Hcd (n=4) infections. RTqPCR was positive in 43/44 patients (97.7% sensitivity), in at least one specimen. Nine out of 863 cases (99% specificity) were RTqPCR positive and therefore classified as possible cases. RTqPCR was positive in 13/30 (43.3%) blood tested in proven cases. A positive RTqPCR in blood was significantly associated with Hcc progressive disseminated histoplasmosis with a positive RTqPCR in 92.3% of the immunocompromised patients with disseminated disease. This new Histoplasma RTqPCR assay enabling amplification of hcc and hcd is highly sensitive and allows the diagnosis of histoplasmosis advantageously from blood and BAL.
Databáze: OpenAIRE
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