Abstract
Massive DNA sequencing studies have expanded our insights
and understanding of the ecological and functional
characteristics of the gut microbiome. Advanced sequencing
technologies allow us to understand the close association of
the gut microbiome with human health and critical illnesses.
In the future, analyses of the gut microbiome will provide
key information associating with human individual health,
which will help provide personalized health care for diseases.
Numerous molecular biological analysis tools have been rapidly
developed and employed for the gut microbiome researches;
however, methodological differences among researchers
lead to inconsistent data, limiting extensive share of
data. It is therefore very essential to standardize the current
method
ologies and establish appropriate pipelines for human
gut microbiome research. Herein, we review the methods
and procedures currently available for studying the human
gut microbiome, including fecal sample collection, metagenomic
DNA extraction, massive DNA sequencing, and data
analyses with bioinformatics. We believe that this review will
contribute to the progress of gut microbiome research in
the clinical and practical aspects of human health.
Citations
Citations to this article as recorded by

- Efficacy of a Postbiotic Formulation Combined With Microneedling for Mild‐to‐Moderate Acne: A Self‐Control Study
Zhanhong Li, Peihui Li, Yu Xu, Changqing Yan, Xiufen Ma, Huiying Wang, Hong Cheng, Jing Zeng, Ting Li, Xinxian Li, Jia Zhou, Jie Zhang, Jianfeng Zhou, Rongya Yang, Yan Wu, Li Li, Wei Lai, Jiangyun Zhao, Zhe Liu, Qiong Meng
Journal of Cosmetic Dermatology.2025;[Epub] CrossRef - Changes in the Gastrointestinal Microbiota Induced by Proton Pump Inhibitors—A Review of Findings from Experimental Trials
Reidar Fossmark, Maya Olaisen
Microorganisms.2024; 12(6): 1110. CrossRef - Deciphering the gut microbiome: The revolution of artificial intelligence in microbiota analysis and intervention
Mohammad Abavisani, Alireza Khoshrou, Sobhan Karbas Foroushan, Negar Ebadpour, Amirhossein Sahebkar
Current Research in Biotechnology.2024; 7: 100211. CrossRef - From the Gut to the Brain: Is Microbiota a New Paradigm in Parkinson’s Disease Treatment?
Cristiana Vilela, Bruna Araújo, Carla Soares-Guedes, Rita Caridade-Silva, Joana Martins-Macedo, Catarina Teixeira, Eduardo D. Gomes, Cristina Prudêncio, Mónica Vieira, Fábio G. Teixeira
Cells.2024; 13(9): 770. CrossRef - Gut Microbiome in Alzheimer’s Disease: from Mice to Humans
Chang Liang, Resel Pereira, Yan Zhang, Olga L. Rojas
Current Neuropharmacology.2024; 22(14): 2314. CrossRef - Comparative Analysis of Healthy Gut Microbiota in German and Korean Populations: Insights from Large-Scale Cohort Studies
Min Kee Son, Yuri Song, Jin Chung, Hee Sam Na
Microbiology Research.2023; 15(1): 109. CrossRef - Changes in the microbiota of the upper airways and intestine in the treatment of acute respiratory infections in preschool children
Tatiana I. Kurdyukova, Olga N. Krasnorutskaya, Daniil Yu. Bugrimov, Artem N. Shevtsov
Russian Pediatric Journal.2023; 23(2): 95. CrossRef - Factors Affecting Gut Microbiota of Puppies from Birth to Weaning
Fatemeh Balouei, Bruno Stefanon, Sandy Sgorlon, Misa Sandri
Animals.2023; 13(4): 578. CrossRef - Culturing the Human Oral Microbiota, Updating Methodologies and Cultivation Techniques
Saber Khelaifia, Pilliol Virginie, Souad Belkacemi, Herve Tassery, Elodie Terrer, Gérard Aboudharam
Microorganisms.2023; 11(4): 836. CrossRef - Influence of Microbiota on Vaccine Effectiveness: “Is the Microbiota the Key to Vaccine-induced Responses?”
So-Hee Hong
Journal of Microbiology.2023; 61(5): 483. CrossRef - gutMDisorder v2.0: a comprehensive database for dysbiosis of gut microbiota in phenotypes and interventions
Changlu Qi, Yiting Cai, Kai Qian, Xuefeng Li, Jialiang Ren, Ping Wang, Tongze Fu, Tianyi Zhao, Liang Cheng, Lei Shi, Xue Zhang
Nucleic Acids Research.2023; 51(D1): D717. CrossRef - The impact of environmental toxins on the animal gut microbiome and their potential to contribute to disease
Dinesh Nalage, Tejswini Sontakke, Ashwini Biradar, Vinod Jogdand, Ravindra Kale, Sanjay Harke, Rahul Kale, Prashant Dixit
Food Chemistry Advances.2023; 3: 100497. CrossRef - Fecal and cloacal microbiomes of cold-stunned loggerhead Caretta caretta, Kemp’s ridley Lepidochelys kempii, and green sea turtles Chelonia mydas
ZR Forbes, AK Scro, SH Patel, KM Dourdeville, RL Prescott, RM Smolowitz
Endangered Species Research.2023; 50: 93. CrossRef - A Two-Step Single Plex PCR Method for Evaluating Key Colonic Microbiota Markers in Young Mexicans with Autism Spectrum Disorders: Protocol and Pilot Epidemiological Application
Julián Herrera-Mejía, Rocío Campos-Vega, Abraham Wall-Medrano, Florinda Jiménez-Vega
Diagnostics.2023; 13(14): 2387. CrossRef - Effect of Concurrent Training on Body Composition and Gut Microbiota in Postmenopausal Women with Overweight or Obesity
MARINE DUPUIT, MÉLANIE RANCE, CLAIRE MOREL, PATRICE BOUILLON, AUDREY BOSCARO, VINCENT MARTIN, EMILIE VAZEILLE, NICOLAS BARNICH, BENOIT CHASSAING, NATHALIE BOISSEAU
Medicine & Science in Sports & Exercise.2022; 54(3): 517. CrossRef - Using integrated meta-omics to appreciate the role of the gut microbiota in epilepsy
Lucia Boeri, Francesca Donnaloja, Marzia Campanile, Lorenzo Sardelli, Marta Tunesi, Federica Fusco, Carmen Giordano, Diego Albani
Neurobiology of Disease.2022; 164: 105614. CrossRef - The gut microbiota as a biomarker in epilepsy
Emilio Russo
Neurobiology of Disease.2022; 163: 105598. CrossRef - Developing whole cell standards for the microbiome field
Chrysi Sergaki, Saba Anwar, Martin Fritzsche, Ryan Mate, Robert J. Francis, Kirsty MacLellan-Gibson, Alastair Logan, Gregory C. A. Amos
Microbiome.2022;[Epub] CrossRef - Changes in Vaginal Microbiome Diversity in Women With Polycystic Ovary Syndrome
Chaoyi Lu, Hui Wang, Jihong Yang, Xinyue Zhang, Yao Chen, Ruizhi Feng, Yun Qian
Frontiers in Cellular and Infection Microbiology.2021;[Epub] CrossRef - High-throughput 16S rDNA sequencing assisting in the detection of bacterial pathogen candidates: a fatal case of necrotizing fasciitis in a child
Dong Qu, Dong-Fang Qiao, Michael Klintschar, Zhi Qu, Xia Yue
International Journal of Legal Medicine.2021; 135(2): 399. CrossRef - Standardization of microbiome studies for urolithiasis: an international consensus agreement
Naveen Kachroo, Dirk Lange, Kristina L. Penniston, Joshua Stern, Gregory Tasian, Petar Bajic, Alan J. Wolfe, Mangesh Suryavanshi, Andrea Ticinesi, Tiziana Meschi, Manoj Monga, Aaron W. Miller
Nature Reviews Urology.2021; 18(5): 303. CrossRef - The Mammalian Metaorganism: A Holistic View on How Microbes of All Kingdoms and Niches Shape Local and Systemic Immunity
Solveig Runge, Stephan Patrick Rosshart
Frontiers in Immunology.2021;[Epub] CrossRef - Diet and the Microbiota–Gut–Brain Axis: Sowing the Seeds of Good Mental Health
Kirsten Berding, Klara Vlckova, Wolfgang Marx, Harriet Schellekens, Catherine Stanton, Gerard Clarke, Felice Jacka, Timothy G Dinan, John F Cryan
Advances in Nutrition.2021; 12(4): 1239. CrossRef - Interrogation of the perturbed gut microbiota in gouty arthritis patients through in silico metabolic modeling
Michael A. Henson
Engineering in Life Sciences.2021; 21(7): 489. CrossRef - A comprehensive review for gut microbes: technologies, interventions, metabolites and diseases
Changlu Qi, Ping Wang, Tongze Fu, Minke Lu, Yiting Cai, Xu Chen, Liang Cheng
Briefings in Functional Genomics.2021; 20(1): 42. CrossRef - Metagenomics Approaches to Investigate the Gut Microbiome of COVID-19 Patients
Sofia Sehli, Imane Allali, Rajaa Chahboune, Youssef Bakri, Najib Al Idrissi, Salsabil Hamdi, Chakib Nejjari, Saaïd Amzazi, Hassan Ghazal
Bioinformatics and Biology Insights.2021;[Epub] CrossRef - Potential role of the skin microbiota in Inflammatory skin diseases
Pan Chen, Guangwen He, Jingru Qian, Yi Zhan, Rong Xiao
Journal of Cosmetic Dermatology.2021; 20(2): 400. CrossRef - Current ocular microbiome investigations limit reproducibility and reliability: Critical review and opportunities
Erin M. Scott, Andrew C. Lewin, Marina L. Leis
Veterinary Ophthalmology.2021; 24(1): 4. CrossRef - Mining microbes for mental health: Determining the role of microbial metabolic pathways in human brain health and disease
Simon Spichak, Thomaz F.S. Bastiaanssen, Kirsten Berding, Klara Vlckova, Gerard Clarke, Timothy G. Dinan, John F. Cryan
Neuroscience & Biobehavioral Reviews.2021; 125: 698. CrossRef - Interactions between immune system and the microbiome of skin, blood and gut in pathogenesis of rosacea
Marie Isolde Joura, Alexandra Brunner, Éva Nemes-Nikodém, Miklós Sárdy, Eszter Ostorházi
Acta Microbiologica et Immunologica Hungarica.2021; 68(1): 1. CrossRef - The gut microbiome in drug‐resistant epilepsy
Simela Chatzikonstantinou, Georgia Gioula, Vasilios K. Kimiskidis, Jack McKenna, Ioannis Mavroudis, Dimitrios Kazis
Epilepsia Open.2021; 6(1): 28. CrossRef - Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
Hiroaki Nakanishi, Katsumi Yoneyama, Masaaki Hara, Aya Takada, Kazuyuki Saito
Scientific Reports.2021;[Epub] CrossRef - Effects of Xylo-Oligosaccharide on the Gut Microbiota of Patients With Ulcerative Colitis in Clinical Remission
Zongwei Li, Zhengpeng Li, Liying Zhu, Ning Dai, Gang Sun, Lihua Peng, Xin Wang, Yunsheng Yang
Frontiers in Nutrition.2021;[Epub] CrossRef - Recent advances in modulating the microbiome
Eamonn M.M Quigley, Prianka Gajula
F1000Research.2020; 9: 46. CrossRef - Tools for Analysis of the Microbiome
Jessica Galloway-Peña, Blake Hanson
Digestive Diseases and Sciences.2020; 65(3): 674. CrossRef - The potential role of interventions impacting on gut-microbiota in epilepsy
Luigi F Iannone, Maria Gómez-Eguílaz, Rita Citaro, Emilio Russo
Expert Review of Clinical Pharmacology.2020; 13(4): 423. CrossRef - State of the Art in the Culture of the Human Microbiota: New Interests and Strategies
Maryam Tidjani Alou, Sabrina Naud, Saber Khelaifia, Marion Bonnet, Jean-Christophe Lagier, Didier Raoult
Clinical Microbiology Reviews.2020;[Epub] CrossRef - Database limitations for studying the human gut microbiome
Camila K Dias, Robert Starke, Victor S. Pylro, Daniel K. Morais
PeerJ Computer Science.2020; 6: e289. CrossRef - Machine learning methods for microbiome studies
Junghyun Namkung
Journal of Microbiology.2020; 58(3): 206. CrossRef - Gut Microbiome Reveals Specific Dysbiosis in Primary Osteoporosis
Zhiming Xu, Zheng Xie, Jianguang Sun, Shilei Huang, Yangyang Chen, Chengxiang Li, Xing Sun, Bicheng Xia, Lingyang Tian, Caijuan Guo, Feng Li, Guofu Pi
Frontiers in Cellular and Infection Microbiology.2020;[Epub] CrossRef - Evaluation of cutaneous, oral and intestinal microbiota in patients affected by pemphigus and bullous pemphigoid: A pilot study
Giovanni Luca Scaglione, Luca Fania, Elisa De Paolis, Maria De Bonis, Cinzia Mazzanti, Giovanni Di Zenzo, Stefania Lechiancole, Serena Messinese, Ettore Capoluongo
Experimental and Molecular Pathology.2020; 112: 104331. CrossRef - Microecology research: a new target for the prevention of asthma
Hong-Lei Shi, Yu-Hao Lan, Zheng-Chuan Hu, Zi-Ning Yan, Ze-Zhong Liu, Xiriaili Kadier, Li Ma, Jin-Yan Yu, Jing Liu
Chinese Medical Journal.2020; 133(22): 2712. CrossRef - Omics Biomarkers in Obesity: Novel Etiological Insights and Targets for Precision Prevention
Krasimira Aleksandrova, Caue Egea Rodrigues, Anna Floegel, Wolfgang Ahrens
Current Obesity Reports.2020; 9(3): 219. CrossRef - Microbiome and colorectal cancer: Roles in carcinogenesis and clinical potential
Ester Saus, Susana Iraola-Guzmán, Jesse R. Willis, Anna Brunet-Vega, Toni Gabaldón
Molecular Aspects of Medicine.2019; 69: 93. CrossRef - Dietary quality and the colonic mucosa–associated gut microbiome in humans
Yanhong Liu, Nadim J Ajami, Hashem B El-Serag, Clark Hair, David Y Graham, Donna L White, Liang Chen, Zhensheng Wang, Sarah Plew, Jennifer Kramer, Rhonda Cole, Ruben Hernaez, Jason Hou, Nisreen Husain, Maria E Jarbrink-Sehgal, Fasiha Kanwal, Gyanprakash K
The American Journal of Clinical Nutrition.2019; 110(3): 701. CrossRef