Validation of a Droplet Digital PCR (ddPCR) Assay to Detect Cyanobacterial 16S rDNA in Human Lung Tissue

Rachael E Barney; Guohong Huang; Torrey L Gallagher; Maeve Tischbein; John DeWitt; Rachel Martindale; Ethan M P LaRochelle; Gregory J Tsongalis; Elijah W Stommel

doi:10.3390/toxics11060531

Validation of a Droplet Digital PCR (ddPCR) Assay to Detect Cyanobacterial 16S rDNA in Human Lung Tissue

Toxics. 2023 Jun 14;11(6):531. doi: 10.3390/toxics11060531.

Authors

Rachael E Barney¹, Guohong Huang¹, Torrey L Gallagher¹, Maeve Tischbein², John DeWitt³, Rachel Martindale³, Ethan M P LaRochelle¹, Gregory J Tsongalis¹, Elijah W Stommel²

Affiliations

¹ Dartmouth-Hitchcock Medical Center, Department of Pathology, Geisel School of Medicine at Dartmouth, Lebanon, NH 03756, USA.
² Dartmouth-Hitchcock Medical Center, Department of Neurology, Geisel School of Medicine at Dartmouth, Lebanon, NH 03756, USA.
³ Department of Pathology, University of Vermont, Burlington, VT 05405, USA.

Abstract

Cyanobacteria produce a variety of secondary metabolites, including toxins that may contribute to the development of disease. Previous work was able to detect the presence of a cyanobacterial marker in human nasal and broncoalveolar lavage samples; however, it was not able to determine the quantification of the marker. To further research the relationship between cyanobacteria and human health, we validated a droplet digital polymerase chain reaction (ddPCR) assay to simultaneously detect the cyanobacterial 16S marker and a human housekeeping gene in human lung tissue samples. The ability to detect cyanobacteria in human samples will allow further research into the role cyanobacteria plays in human health and disease.

Keywords: 16S; ALS; cyanobacteria; ddPCR; human tissue.

Grants and funding

This research was funded by ALS Association, 18-IIA-437 Stommel (PI) 01/01/2018–04/30/2021, Investigating aerosolized cyanobacteria in amyotrophic lateral sclerosis.