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

A Alanio; D Garcia-Hermoso; S Mercier-Delarue; F Lanternier; M Gits-Muselli; J Menotti; B Denis; A Bergeron; M Legrand; O Lortholary; S Bretagne; French Mycosis Study Group

doi:10.1016/j.cmi.2015.01.017

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

Clin Microbiol Infect. 2015 Jun;21(6):594.e1-5. doi: 10.1016/j.cmi.2015.01.017. Epub 2015 Jan 28.

Authors

A Alanio¹, D Garcia-Hermoso², S Mercier-Delarue³, F Lanternier⁴, M Gits-Muselli⁵, J Menotti⁶, B Denis⁷, A Bergeron⁸, M Legrand⁹, O Lortholary¹⁰, S Bretagne¹¹; French Mycosis Study Group

Collaborators

French Mycosis Study Group:
J Maizel, T Chouaki, M Hunault, J P Bouchara, M Sirodot, J Gaillat, X Blanc, Q Gobert, J Leroy, F Legrand, J Navallou, F Grenouillet, L Million, G Blasco, D Malvy, I Accoceberry, S Ansart, E Moalic, O Reman, R Verdon, J Bonhomme, D Caillot, P Charles, P Chavanet, F Dalle, J Stahl, O Epaulard, A Thiebaut-Bertrand, M Cornet, B Guery, B Sendid, T Ferry, T Baudry, M Michallet, M Moutran, P Miailhes, A Bienvenue, C Faucher, S Gayet, S Ranque, O Jonquet, P Moreau, F Morio, P Roger, P Dellamonica, M Gari-Toussaint, A Veinstein, J Saulnier, C Kauffmann, C Arvieux, M Bernard, V Gandemer, J P Gangneux, C Joly, G Pfister, B Veber, L Favennec, C Cazorla, H Raberin, M Alvarez, A Huynh, S Cassaing, E Giroux, J Dunand, J Naccache, C Gardin, O Freynet, S Brun, E Azoulay, C Lacroix, E Raffoux, V Joly, C Chochillon, J Charpentier, A Paugam, E Rivaud, P Cahen, A Novara, E Danaoui, C Cordonnier, F Botterel, S Bretagne, N Anguel, A Angoulvant, F Suarez, F Lacaille, M Oualha, M Bougnoux, A Combes, P Buffet, B Clair, J Dunand, E Maury, C Hennequin, M Tabone, B Wyplosz, M Merad, L Mihaila

Affiliations

¹ Laboratoire de Parasitologie-Mycologie, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, France; Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Paris, France. Electronic address: [email protected].
² Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Paris, France.
³ Laboratoire de Microbiologie, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France.
⁴ Université Paris Descartes, Hôpital Necker-Enfants malades, Assistance Publique-Hôpitaux de Paris (APHP), Centre d'Infectiologie Necker-Pasteur, Institut Imagine, Paris, France; Université Paris Descartes, Paris, France.
⁵ Laboratoire de Parasitologie-Mycologie, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France.
⁶ Laboratoire de Parasitologie-Mycologie, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, France; Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France.
⁷ Service de Maladies Infectieuses et Tropicales, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France.
⁸ Université Paris Diderot, Sorbonne Paris Cité, Paris, France; UMR-S717 INSERM, Service de Pneumologie, AP-HP, Hôpital Saint Louis, Paris, France.
⁹ Université Paris Diderot, Sorbonne Paris Cité, Paris, France; UMR 942, INSERM, Service d'Anesthésie-Réanimation, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France.
¹⁰ Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Paris, France; Université Paris Descartes, Hôpital Necker-Enfants malades, Assistance Publique-Hôpitaux de Paris (APHP), Centre d'Infectiologie Necker-Pasteur, Institut Imagine, Paris, France; Université Paris Descartes, Paris, France.
¹¹ Laboratoire de Parasitologie-Mycologie, AP-HP, Groupe Hospitalier Saint-Louis-Lariboisière-Fernand-Widal, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, France; Institut Pasteur, Unité de Mycologie Moléculaire, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France; CNRS URA3012, Paris, France.

PMID: 25726039
DOI: 10.1016/j.cmi.2015.01.017

Abstract

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; Mucorales identification; PCR; electrospray-ionization mass spectrometry; mucormycosis; species level.

Publication types

Comparative Study
Evaluation Study

MeSH terms

DNA, Fungal / chemistry
DNA, Fungal / genetics
Humans
Microbiological Techniques / methods
Mucorales / genetics
Mucorales / isolation & purification*
Mucormycosis / diagnosis*
Pathology, Molecular / methods*
Polymerase Chain Reaction / methods*
Prospective Studies
Real-Time Polymerase Chain Reaction / methods
Sequence Analysis, DNA / methods
Spectrometry, Mass, Electrospray Ionization / methods*
Time Factors

Substances

DNA, Fungal