Rapid assessment of viral water quality using a novel recombinase polymerase amplification test for human adenovirus
Sensitive and rapid methods for determining viral contamination of water are critical, since illness can be caused by low numbers of viruses and bacterial indicators do not adequately predict viral loads. We developed novel rapid assays for detecting the viral water quality indicator human adenovirus (HAdV). A simple 15-min recombinase polymerase amplification step followed by a 5-min lateral flow detection is used. Species-specific assays were developed to discriminate HAdV A, B, C and F, and combined into a multiplex test (Ad-FAC). Species-specific assays enabled detection of 10–50 copies of the HAdV plasmid. Sample testing using methods optimised for wastewater analysis indicated the Ad-FAC assay showed 100% sensitivity and 100% specificity when compared with HAdV qPCR, with a detection limit as low as 50 gene copies. This is the first study to demonstrate the use of RPA for detecting enteric viruses in water samples, to assess virological water quality. The ability to rapidly detect enteric virus contamination of water could assist in more effective management of water safety and better protection of public health.
KeywordsRecombinase polymerase amplification Lateral flow immunoassay Water quality Adenovirus Inhibition Isothermal amplification
The authors thank Maxim Scheludchenko and Anna Padovan for collection of wastewater samples and PEG precipitations. We also thank Professor Richard Burns for critical review of the manuscript, and we are grateful to Helen Stratton, Anne Roiko and Charles Lemckert for advice and encouragement.
This study was funded by a University of the Sunshine Coast (USC, Australia) Faculty Seed Grant. Author1 received a scholarship provided by USC and Griffith University as part of a PONDS Project funded by the Queensland Government DSITIA Science Fund project, through the Smart Water Research Centre (Griffith University, Australia).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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