Sequence-based Methods for Detecting and Evaluating the Human Gut Mycobiome

Authors

Mallory J. Suhr, University of Nebraska-LincolnFollow
Nabaraj Banjara, University of Nebraska-LincolnFollow
Heather E. Hallen-Adams, University of Nebraska-LincolnFollow

Document Type

Article

Date of this Version

2016

Citation

Suhr MJ, Banjara N, Hallen-Adams HE (2016) Sequence-based methods for detecting and evaluating the human gut mycobiome. Lett Appl Microbiol 62:209-215.
doi:10.1111/lam.12539

Comments

Copyright © 2015 The Society for Applied Microbiology; published by Wiley. Used by permission.

Abstract

We surveyed the fungal microbiota in 16 fecal samples from healthy humans with a vegetarian diet. Fungi were identified using molecular cloning, 454 pyrosequencing and a Luminex analyte specific reagent (ASR) assay, all targeting the ITS region of the rRNA genes. Fungi were detected in each fecal sample and at least 46 distinct fungal operational taxonomic units (OTUs) were detected, from two phyla — Ascomycota and Basidiomycota. Fusarium was the most abundant genus, followed by Malassezia, Penicillium, Aspergillus, and Candida. Commonly detected fungi such as Aspergillus and Penicillium, as well as known dietary fungi Agaricus bisporus and Ophiocordyceps sinensis, are presumed to be transient, allochthonous members due to their abundance in the environment or dietary associations. No single method identified the full diversity of fungi in all samples; pyrosequencing detected more distinct OTUs than the other methods, but failed to detect OTUs in some samples that were detected by cloning and/or ASR assays. ASRs were limited by the commercially available assays, but the potential to design new, optimized assays, coupled with speed and cost, makes the ASR method worthy of further study.