A multiplexed droplet digital PCR assay performs better than qPCR on inhibition prone samples

Ruth Hall Sedlak; Jane Kuypers; Keith R Jerome

doi:10.1016/j.diagmicrobio.2014.09.004

A multiplexed droplet digital PCR assay performs better than qPCR on inhibition prone samples

Diagn Microbiol Infect Dis. 2014 Dec;80(4):285-6. doi: 10.1016/j.diagmicrobio.2014.09.004. Epub 2014 Sep 16.

Authors

Ruth Hall Sedlak¹, Jane Kuypers¹, Keith R Jerome²

Affiliations

¹ Molecular Virology Laboratory, Department of Laboratory Medicine, University of Washington, Seattle, WA.
² Molecular Virology Laboratory, Department of Laboratory Medicine, University of Washington, Seattle, WA; Vaccine Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA. Electronic address: kjerome@fhcrc.org.

PMID: 25277746
DOI: 10.1016/j.diagmicrobio.2014.09.004

Abstract

We demonstrate the development of a multiplex droplet digital PCR assay for human cytomegalovirus (CMV), human adenovirus species F, and an internal plasmid control that may be useful for PCR inhibition-prone clinical samples. This assay performs better on inhibition-prone stool samples than a quantitative PCR assay for CMV and is the first published clinical virology droplet digital PCR assay to incorporate an internal control.

Keywords: Digital PCR; Inhibition; Quantitative PCR; Stool.

Publication types

Comparative Study

MeSH terms

Adenoviruses, Human / genetics*
Cytomegalovirus / genetics*
Cytomegalovirus Infections / virology
Feces / virology*
Humans
Multiplex Polymerase Chain Reaction / methods*
Real-Time Polymerase Chain Reaction