---

About Next Generation 16S Sequencing

Many sites of the human body are colonized by complex communities of microbes (the "human microbiome") in both health and various disease states. Highly diverse, polymicrobial specimens are often difficult, or even impossible, to fully characterize by techniques in common clinical use:

• Culture-based identification relies upon the ability of organisms to grow and replicate in vitro. Therefore, detection of fastidious or slow-growing organisms, or those rendered inviable due to processing (such as in formalin-fixed paraffin embedded tissue specimens) or during storage (such as anaerobes which have been exposed to oxygen) is limited. Furthermore, only a limited number of species can be practically classified by this approach.
• Molecular methods, such as our 16S rRNA gene sequencing assay, do not require prior culture and have improved ability to identify inviable or fastidious organisms from direct specimens. However, in a polymicrobial speciemen only the predominant organism may be identified, or an uninterpretable signal may be generated (Figure 1).

In contrast to conventional approaches, next-generation DNA sequencing (alternatively termed "NGS", "high-throughput sequencing", "massively parallel sequencing", or "deep sequencing") provides independent sequence data from millions of individual DNA molecules (Figure 2), allowing each fragment to be classified independently.

This unique ability extends upon the advantages of current molecular methods by allowing us to catalog the organisms present within even very complex polymicrobial bacterial communities, directly from patient specimens.

---

Information on Available Assays

Our lab currently offers high-fidelity Illumina next-generation DNA sequencing of clinical specimens which contain multiple bacterial DNA templates. Methods are validated for the purpose of clinical molecular diagnosis and patient care. Research services are also available - please contact us for additional information.

This test is available as reflex testing for specimens which are expected to be polymicrobial based on broad range bacterial PCR.

To learn more about clinical applications and methodology, please see the following publications from our lab:

• Co-infection of Fusobacterium nucleatum and Actinomyces israelii in mastoiditis diagnosed by next-generation DNA sequencing
• Molecular diagnosis of Actinomadura madurae infection by 16S rRNA deep sequencing

---

---

1. Home
2. Available Tests
3. 16S Next Generation Sequencing