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Evaluation of a New Histoplasma spp. Quantitative RT-PCR Assay

2021; Elsevier BV; Volume: 23; Issue: 6 Linguagem: Inglês

10.1016/j.jmoldx.2021.02.007

1943-7811

Alexandre Alanio, Maud Gits‐Muselli, Fanny Lanternier, Aude Sturny-Leclère, Marion Benazra, Samia Hamane, Anderson Messias Rodrigues, Dea Garcia‐Hermoso, Olivier Lortholary, Françoise Dromer, Stéphane Bretagne, M. Woimant, Geoffrey L. Smith, Souad Silhadi, Nicolas Vignier, A. Pitsch, K. Jidar, Nicolas Traversier, D.M. Poisson, Claire Lecointre, F. Foulet, Françoise Botterel, Nawel Ait Ammar, Amsellem, Frédéric Gabriel, Philipe Poirier, Marjorie Cornu, Séverine Loridant, Florent Morio, D. Boutoille, Fakhri Jeddi, Lilia Hasseine, Rachida Ouissa, D. Toubas, Éric Bailly, Guillaume Désoubeaux, Emily Ronez, G Foulon, Sebastien Lefrançois, Christine Bonnal, A. Paugam, Maxime Dougados, Marine Desroches, H. Barazzutti, N. Paleiron, Méja Rabodonirina, Émilie Catherinot, Émilie Cardot-Martin, Chrisian Hiesse, Hélène Salvator, Claire Aguilar, Anne Gigandon, Thomas de Montpreville, Marie‐Elisabeth Bougnoux, Emilie Sitterlé, Arnaud Fekkar, Sébastien Imbert, Alexandre Bleibtreu, Yaye Senghor, Blandine Denis, Jean‐Michel Molina, Geoffroy Liégeon, Anne-Lise Munnier, Marion Malphettes, Julie Denis, Alain Berlioz-Arthaud, Franciska Lange, Myriam Chiaruzzi, Loïc Epelboin,

Plant Pathogens and Fungal Diseases

Laboratory diagnosis of histoplasmosis is based on various methods, including microscopy, culture, antigen, and DNA detection of Histoplasma capsulatum var. capsulatum or Histoplasma capsulatum var. duboisii. To improve sensitivity of existing real-time quantitative PCR (qPCR) assays, we developed a new RT-qPCR assay that allows amplification of whole nucleic acids of Histoplasma spp. 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 1319 prospectively collected consecutive samples from 907 patients suspected of having histoplasmosis were tested routinely between May 2015 and May 2019 in parallel with standard diagnostic procedures performed in parallel. Forty-four had proven histoplasmosis attributable to H. capsulatum var. capsulatum (n = 40) or H. capsulatum var. duboisii (n = 4) infections. The results of RT-qPCR were positive in 43 of 44 patients (97.7% sensitivity) in at least one specimen. Nine of 863 cases (99% specificity) were RT-qPCR positive and therefore classified as possible cases. RT-qPCR was positive in 13 of 30 (43.3%) blood samples tested in proven cases. A positive RT-qPCR result in blood was significantly associated with H. capsulatum var. capsulatum progressively disseminated histoplasmosis with a positive RT-qPCR result in 92.3% of the immunocompromised patients with disseminated disease. This new Histoplasma RT-qPCR assay enabling amplification of H. capsulatum var. capsulatum and H. capsulatum var. duboisii is highly sensitive and allows the diagnosis of histoplasmosis advantageously from blood and bronchoalveolar lavage fluid.