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Field-based evaluation of a male-specific (F+) RNA coliphage concentration method
J C Chandler1, A Pérez-Méndez2, J Paar3
1National Wildlife Research Center, Wildlife Services, Animal and Plant Health Inspection Service, United States Department of Agriculture, Fort Collins, CO, USA.
Journal of Virological Methods
|November 7, 2016
Summary
A new method using anion exchange resin effectively concentrates fecal indicator viruses from environmental water. This technique aids in detecting fecal contamination and assessing water safety for public health.
Area of Science:
- Environmental microbiology
- Public health science
- Molecular biology
Background:
- Fecal contamination of water is a major public health concern due to enteric virus presence.
- Sensitive, reliable, and user-friendly methods are crucial for monitoring water quality and safety.
- Male-specific (F+) RNA coliphages (FRNA) serve as key fecal indicator organisms.
Purpose of the Study:
- To evaluate an anion exchange resin-based method for concentrating FRNA coliphages from diverse environmental waters.
- To assess the method's effectiveness in detecting fecal contamination in water suspected of being feces-contaminated.
- To investigate matrix-dependent inhibition affecting real-time reverse transcriptase (RT)-PCR detection.
Main Methods:
- Field evaluation of an anion exchange resin method for FRNA coliphage concentration.
- Direct nucleic acid isolation from adsorbed coliphages.
- Real-time reverse transcriptase (RT)-PCR for FRNA coliphage detection and quantification.
- Evaluation of matrix-dependent inhibition using spiked FRNA coliphages from four genogroups (GI-GIV).
Main Results:
- RT-PCR detection success rates for spiked FRNA coliphages were high: 97% (GI), 72% (GII), 85% (GIII), and 98% (GIV).
- Inhibitors in water samples affected RT-PCR assay efficiency, with no correlation to physicochemical properties.
- Naturally occurring FRNA coliphages were detected in 61.5% (40 of 65) of environmental water samples.
Conclusions:
- The anion exchange resin method is a viable system for concentrating FRNA coliphages from environmental waters.
- The method demonstrates potential for routine monitoring of fecal contamination in water sources.
- Understanding and mitigating RT-PCR inhibition is important for accurate water quality assessment.
