Related Experiment Video
Updated: Mar 21, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Intricacies of assessing the human microbiome in epidemiologic studies
Courtney K Robinson1, Rebecca M Brotman2, Jacques Ravel3
1Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore.
Purpose:
In the past decade, remarkable relationships have been documented between dysbiosis of the human microbiota and adverse health outcomes. This review seeks to highlight some of the challenges and pitfalls that may be encountered during all stages of microbiota research, from study design and sample collection, to nucleic acid extraction and sequencing, and bioinformatic and statistical analysis.
Methods:
Literature focused on human microbiota research was reviewed and summarized.
Results:
Although most studies have focused on surveying the composition of the microbiota, fewer have explored the causal roles of these bacteria, archaea, viruses, and fungi in affecting disease states. Microbiome research is in its relatively early years and many aspects remain challenging, including the complexity and personalized aspects of microbial communities, the influence of exogenous and often confounding factors, the need to apply fundamental principles of ecology and epidemiology, the necessity for new software tools, and the rapidly evolving genomic, technological, and analytical landscapes.
Conclusions:
Incorporating human microbiome research in large epidemiologic studies will soon help us unravel the intricate relationships that we have with our microbial partners and provide interventional opportunities to improve human health.
Related Concept Videos
Introduction to the Human Microbiota
Methods to Assess Microbial Communities
Development of Human Microbiota
Methods to Assess Microbial Populations
The Oral Microbiota
Human Virome

