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Modern Molecular Taxonomy01:29

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Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
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Related Experiment Video

Updated: Jan 1, 2026

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
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Microbiome Diagnostics.

Robert Schlaberg1,2,3

  • 1Department of Pathology, University of Utah, Salt Lake City, UT.

Clinical Chemistry
|December 18, 2019
PubMed
Summary

Advances in sequencing and computational biology enable human microbiota profiling for diagnostics and therapeutics. Standardization and interdisciplinary collaboration are key to implementing these microbiome insights into routine medical practice.

Area of Science:

  • Microbiology
  • Genomics
  • Computational Biology

Background:

  • Recent advances in sequencing technology and computational biology have illuminated the significant role of the human microbiota in health and disease.
  • There is a growing interest across medical specialties in utilizing accurate and reproducible microbiota profiling for diagnostic and therapeutic purposes.

Purpose of the Study:

  • To review current approaches, challenges, and considerations for diagnostic applications of microbiota profiling.
  • To discuss methodological advancements from 16S rRNA sequencing to metagenomics and metatranscriptomics.
  • To highlight the need for standardization and quality control in microbiome testing.

Main Methods:

  • Review of existing literature on microbiota profiling techniques.

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  • Discussion of various sequencing-based approaches including amplicon sequencing, metagenomics, and metatranscriptomics.
  • Analysis of challenges in reagent purity, reference databases, and data analysis solutions.

    • Main Results:

    • The body of evidence linking microbiota to human health and disease is rapidly expanding.
    • Therapeutic manipulation of the intestinal microbiota is becoming a standard practice.
    • Technological and scientific advancements are paving the way for microbiome diagnostics.

    Conclusions:

    • Standardization of vulnerable testing workflow steps and QC standards are crucial for improving replicability.
    • Addressing challenges in reagent purity, database quality, and data analysis requires interdisciplinary collaboration.
    • The next decade is expected to see the integration of microbiome diagnostics into routine clinical practice.