Metagenomic detection of phage-encoded platelet-binding factors in the human oral cavity

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 108; no. Supplement 1; pp. 4547 - 4553
• Main Authors: Willner, Dana, Furlan, Mike, Schmieder, Robert, Grasis, Juris A, Pride, David T, Relman, David A, Angly, Florent E, McDole, Tracey, Mariella, Ray P. Jr, Rohwer, Forest, Haynes, Matthew
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 15.03.2011; National Acad Sciences
• Series: Colloquium Paper
• Subjects: Bacteriophages - genetics; Bacteriophages - isolation & purification; Base Sequence; Biological Sciences; Blood Platelets - metabolism; California; Chloroform; coliphages; Computational Biology; Contamination; DNA; Endocarditis; Endocarditis - virology; Escherichia coli; Escherichia coli - virology; Flow Cytometry; genes; Genes, Viral - genetics; Genomes; heart; Humans; Metagenomics; microbial contamination; microorganisms; Molecular Sequence Data; mouth; Mouth - microbiology; Mouth - virology; Nucleotide sequence; Oral cavity; Oropharynx; Pathogens; Phage T3; Phages; Phylogeny; Platelets; Polymerase chain reaction; Propionibacterium acnes; Propionibacterium acnes - virology; rapid methods; risk; Saliva; Sequence Analysis, DNA; Streptococcus; Streptococcus mitis; Streptococcus mitis - virology; Virulence; California
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1000089107

The human oropharynx is a reservoir for many potential pathogens, including streptococcal species that cause endocarditis. Although oropharyngeal microbes have been well described, viral communities are essentially uncharacterized. We conducted a metagenomic study to determine the composition of oropharyngeal DNA viral communities (both phage and eukaryotic viruses) in healthy individuals and to evaluate oropharyngeal swabs as a rapid method for viral detection. Viral DNA was extracted from 19 pooled oropharyngeal swabs and sequenced. Viral communities consisted almost exclusively of phage, and complete genomes of several phage were recovered, including Escherichia coli phage T3, Propionibacterium acnes phage PA6, and Streptococcus mitis phage SM1. Phage relative abundances changed dramatically depending on whether samples were chloroform treated or filtered to remove microbial contamination. pblA and pblB genes of phage SM1 were detected in the metagenomes. pblA and pblB mediate the attachment of S. mitis to platelets and play a significant role in S. mitis virulence in the endocardium, but have never previously been detected in the oral cavity. These genes were also identified in salivary metagenomes from three individuals at three time points and in individual saliva samples by PCR. Additionally, we demonstrate that phage SM1 can be induced by commonly ingested substances. Our results indicate that the oral cavity is a reservoir for pblA and pblB genes and for phage SM1 itself. Further studies will determine the association between pblA and pblB genes in the oral cavity and the risk of endocarditis.