Revisiting the Fungal ITS2 Sequencing Data from the Human Microbiome Project

Abstract While the Human Microbiome Project (HMP) significantly advanced our understanding of the human microbiome, our revisiting of its fecal ITS2 sequencing data reveals two significant issues. First, technical variations in sequencing or data processing introduce batch effects in downstream analyses. Second, we discovered a mismatch between the reported and actual amplification primers, which requires careful consideration when interpreting, reusing, or referencing HMP ITS2 data and methodology. Competing Interest Statement The authors have declared no competing interest. Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.