The development of high quality mock communities for microbiome analysis as a positive control

Abstract

It has been almost 20 years since the advent of Next-Generation Sequencing (NGS), which enabled a huge expansion in our understanding of microbiology. However, issues regarding the accuracy and experimental reliability of NGS for microbiota remain. This is primarily because NGS analysis involves multiple processes and methods, each of which introduces experimental biases. Identifying these experimental biases and optimizing experimental methods is an ongoing process to improve the reliability of NGS analysis. To achieve this, reference materials representative of natural microbiota, referred to as Mock Communities (MCs), are required as positive controls. New MCs have been developed for the NGS analysis of human intestinal microbiota and have been available since 2022 from our organization, NITE (National Institute of Technology and Evaluation). To create these high-quality MCs, an optimized method for measuring the biological mass of bacteria was developed. This paper describes the development of these MCs and introduces the diverse MCs currently available worldwide.