Use of F-Specific RNA Bacteriophage to Estimate Infectious Norovirus Levels in Oysters View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-05-21

AUTHORS

J. A. Lowther, L. Cross, T. Stapleton, N. E. Gustar, D. I. Walker, M. Sills, S. Treagus, V. Pollington, D. N. Lees

ABSTRACT

Contamination of bivalve shellfish, particularly oysters, with norovirus is recognised as a significant food safety risk. Methods for quantification of norovirus in oysters using the quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) are well established, and various studies using RT-qPCR have detected norovirus in a considerable proportion of oyster samples, both in the UK and elsewhere. However, RT-qPCR detects viral genome, and by its nature is unable to discriminate between positive results caused by infectious viruses and those caused by non-infectious remnants including damaged virus particles and naked RNA. As a result, a number of alternative or complementary approaches to RT-qPCR testing have been proposed, including the use of infectious viral indicator organisms, most frequently F-specific RNA bacteriophage (F-RNA phage). In this study, we investigated the relationships between F-RNA phage and norovirus in digestive tissues from two sets of oyster samples, one randomly collected at retail (630 samples), and one linked to suspected norovirus illness outbreaks (nine samples). A positive association and correlation between PCR-detectable levels of genogroup II F-RNA bacteriophage (associated with human faecal contamination) and norovirus was found in both sets of samples, with more samples positive for genogroup II phage, at generally higher levels than norovirus. Levels of both viruses were higher in outbreak-related than retail samples. Infectious F-RNA phage was detected in 47.8% of all retail samples, and for a subset of 224 samples where characterisation of phage was carried out, infectious GII phage was detected in 30.4%. Infectious GII phage was detected in all outbreak-related samples. Determination of infectivity ratios by comparing levels of PCR-detectable (copies/g) and infectious GII phage (pfu/g) revealed that in the majority of cases less than 10% of virus detected by RT-qPCR was infectious. Application of these ratios to estimate infectious norovirus levels indicated that while 77.8% of outbreak-related samples contained > 5 estimated infectious norovirus/g, only 13.7% of retail samples did. Use of a combination of levels of PCR-detectable norovirus and infectious F-RNA phage showed that while only 7.0% of retail samples contained both > 100 copies/g norovirus and > 10 pfu/g F-RNA phage, these combined levels were present in 77.8% of outbreak-related samples, and 75.9% of retail samples with > 5 estimated infectious norovirus/g. We therefore suggest that combining RT-qPCR testing with a test for infectious F-RNA phage has the potential to better estimate health risks, and to better predict the presence of infectious norovirus than RT-qPCR testing alone. More... »

PAGES

247-258

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s12560-019-09383-3

DOI

http://dx.doi.org/10.1007/s12560-019-09383-3

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1115024316

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/31115869


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73 infectivity ratio
74 levels
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78 method
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80 naked RNA
81 nature
82 non-infectious remnants
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85 norovirus levels
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91 oyster samples
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95 polymerase chain reaction
96 positive association
97 positive results
98 potential
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100 proportion
101 quantification
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103 quantitative real-time reverse transcription-polymerase chain reaction
104 ratio
105 reaction
106 real-time reverse transcription-polymerase chain reaction
107 relationship
108 remnants
109 results
110 retail
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112 reverse transcription-polymerase chain reaction
113 risk
114 safety risks
115 samples
116 set
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118 shellfish
119 significant food safety risk
120 specific RNA bacteriophages
121 study
122 subset
123 test
124 testing
125 tissue
126 transcription-polymerase chain reaction
127 use
128 viral genome
129 viral indicator organisms
130 virus
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