Abstract

CrAssphage is a recently discovered human gut–associated bacteriophage. To validate the potential use of crAssphage for detecting human fecal contamination on environmental surfaces and hands, we tested stool samples (n = 60), hand samples (n = 30), and environmental swab samples (n = 201) from 17 norovirus outbreaks for crAssphage by real-time PCR. In addition, we tested stool samples from healthy persons (n = 173), respiratory samples (n = 113), and animal fecal specimens (n = 68) and further sequenced positive samples. Overall, we detected crAssphage in 71.4% of outbreak stool samples, 48%–68.5% of stool samples from healthy persons, 56.2% of environmental swabs, and 60% of hand rinse samples, but not in human respiratory samples or animal fecal samples. CrAssphage sequences could be grouped into 2 major genetic clusters. Our data suggest that crAssphage could be used to detect human fecal contamination on environmental surfaces and hands.

Highlights

  • CrAssphage is a recently discovered human gut–associated bacteriophage

  • The high prevalence of crAssphage on surfaces and hands in norovirus outbreak settings suggests that these phages can be used as an indicator to monitor human fecal contamination of environmental sources other than sewage-contaminated water [21,22,23,24,25]

  • CrAssphage contamination was frequently found on environmental surfaces in public areas of cruise ships both during and after norovirus outbreaks, suggesting a potential role of crAssphage in monitoring fecal contamination on surfaces in common settings that could be targeted for enhanced cleaning and disinfection practices

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Summary

Introduction

CrAssphage is a recently discovered human gut–associated bacteriophage. To validate the potential use of crAssphage for detecting human fecal contamination on environmental surfaces and hands, we tested stool samples (n = 60), hand samples (n = 30), and environmental swab samples (n = 201) from 17 norovirus outbreaks for crAssphage by real-time PCR. We detected crAssphage in 71.4% of outbreak stool samples, 48%–68.5% of stool samples from healthy persons, 56.2% of environmental swabs, and 60% of hand rinse samples, but not in human respiratory samples or animal fecal samples. Over the past few decades, several viruses (e.g., human polyomavirus, Aichi virus, norovirus, and human adenovirus) have been studied as human fecal indicators for the detection of sewage-contaminated source and drinking water [10,11,12,13] Both norovirus and adenovirus have been suggested as potential biomarkers of viral contamination to assess hygiene status and potential human health risk of contaminated surfaces and hands of affected persons [4,12,14,15,16,17]. To confirm the specificity of crAssphage for the human gut, we tested fecal specimens from cats, rats, rhesus monkeys, and husbandry animals (cows, pigs, sheep, and horses)

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