Primers matter: Influence of the primer selection on human fungal detection using high throughput sequencing

Gut Microbes. 2022 Jan-Dec;14(1):2110638. doi: 10.1080/19490976.2022.2110638.

Abstract

Microbiota research has received an increasing attention for its role in disease development and fungi are considered as one of the key players in the microbial niche. Various sequencing approaches have been applied to uncover the role of fungal community in health and disease; however, little is known on the performance of various primers and comparability between the studies. Motivated by the recent publications, we performed a systematic comparison of the 18S and ITS regions to identify the impact of various primers on the sequencing results. Using four pairs of primers extensively used in literature, fungal community was retrieve from 25 fecal samples, and applying high throughput sequencing; and the results were compared in order to select the most suitable primers for fungal detection in human fecal samples. Considering the high variability between samples, primers described in the Earth microbiome project detected the broadest fungal spectrum suggesting its superior performance in mycobiome research.

Keywords: 18S; ITS; Mycobiome; fungi; sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Fungal
  • Fungi / genetics
  • Gastrointestinal Microbiome* / genetics
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Mycobiome* / genetics

Substances

  • DNA, Fungal

Grants and funding

The study is partially supported by the funds of European Commission through the “European funds for regional development” (EFRE) as well as by the regional Ministry of Economy, Science and Digitalization of Saxony-Anhalt as part of the “Autonomy in old Age” (AiA) research group for “LiLife” Project to AL (Project ID: ZS/2018/11/95324; European Regional Development Fund.