Droplet digital PCR for simultaneous quantification of general and human-associated fecal indicators for water quality assessment

Despite wide application to beach water monitoring and microbial source identification, results produced by quantitative PCR (qPCR) methods are subject to bias introduced by reliance on quantitative standards. Digital PCR technology provides direct, standards-free quantification and may potentially...

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Bibliographic Details
Published in:Water research (Oxford) 2015-03, Vol.70, p.337-349
Main Authors: Cao, Yiping, Raith, Meredith R., Griffith, John F.
Format: Article
Language:English
Subjects:
Assessments
Bacteroidetes - genetics
Bacteroidetes - isolation & purification
Biomarkers - analysis
Digital
Digital PCR
Droplets
Enterococcus
Enterococcus - genetics
Enterococcus - isolation & purification
Environmental Monitoring - methods
Feces - microbiology
Human fecal-associated Bacteroidales
Humans
Inhibition
Inhibitors
Markers
Microbial source tracking
Monitoring
Real-Time Polymerase Chain Reaction
Recreational water quality
Sensitivity and Specificity
Water Quality
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Summary:Despite wide application to beach water monitoring and microbial source identification, results produced by quantitative PCR (qPCR) methods are subject to bias introduced by reliance on quantitative standards. Digital PCR technology provides direct, standards-free quantification and may potentially alleviate or greatly reduce other qPCR limitations such as difficulty in multiplexing and susceptibility to PCR inhibition. This study examined the efficacy of employing a duplex droplet digital PCR (ddPCR) assay that simultaneously quantifies Enterococcus spp. and the human fecal-associated HF183 marker for water quality assessment. Duplex ddPCR performance was evaluated side-by-side with qPCR and simplex ddPCR using reference material and 131 fecal and water samples. Results for fecal and water samples were highly correlated between ddPCR and simplex qPCR (coefficients > 0.93, p 
ISSN:0043-1354
1879-2448
DOI:10.1016/j.watres.2014.12.008