Molecular identification of Mucorales in human tissues: contribution of PCR electrospray-ionization mass spectrometry

Alexandre Alanio (1, 2, 3, 4) , D. Garcia-Hermoso (3, 4) , S. Mercier-Delarue (5) , F. Lanternier (6, 7) , M. Gits-Muselli (1) , J. Menotti (1, 4) , B. Denis , A. Bergeron (8, 2) , Michel Legrand (9, 2) , O. Lortholary (3, 4, 6, 7) , S. Bretagne (1, 2, 3, 4) , French Mycosis Study Group

1 Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris]
2 UPD7 - Université Paris Diderot - Paris 7
3 Mycologie moléculaire
4 CNRMA - Centre National de Référence des Mycoses invasives et antifongiques - Mycologie moléculaire
5 Laboratoire de microbiologie
6 Centre d'infectiologie Necker-Pasteur [CHU Necker]
7 UPD5 - Université Paris Descartes - Paris 5
8 Service de pneumologie [Saint-Louis]
9 Service d’anesthésie-réanimation [AP-HP Hôpital Saint-Louis]

Molecular methods are crucial for mucormycosis diagnosis because cultures are frequently negative, even if microscopy suggests the presence of hyphae in tissues. We assessed PCR/electrospray-ionization mass spectrometry (PCR/ESI-MS) for Mucorales identification in 19 unfixed tissue samples from 13 patients with proven or probable mucormycosis and compared the results with culture, quantitative real-time PCR, 16S-23S rRNA gene internal transcribed spacer region (ITS PCR) and 18S PCR sequencing. Concordance with culture identification to both genus and species levels was higher for PCR/ESI-MS than for the other techniques. Thus, PCR/ESI-MS is suitable for Mucorales identification, within 6 hours, for tissue samples for which microscopy results suggest the presence of hyphae.

Keywords: Culture; DNA; Fungal; electrospray-ionization mass spectrometry; Humans; Microbiological Techniques; Mucorales; Mucorales identification; Mucormycosis; Pathology; Molecular; PCR; Polymerase Chain Reaction; Prospective Studies; Real-Time Polymerase Chain Reaction; Sequence Analysis; species level; Spectrometry; Mass; Electrospray Ionization; Time Factors

Domains: Life Sciences [q-bio]; Life Sciences [q-bio]; Microbiology and Parasitology; Mycology

Alexandre Alanio, D. Garcia-Hermoso, S. Mercier-Delarue, F. Lanternier, M. Gits-Muselli, et al.. Molecular identification of Mucorales in human tissues: contribution of PCR electrospray-ionization mass spectrometry. Clinical Microbiology and Infection, 2015, 21 (6), pp.594.e1--5. ⟨10.1016/j.cmi.2015.01.017⟩. ⟨pasteur-01405221⟩