Publication: Norovirus Monitoring in Oysters Using Two Different Extraction Methods
Issued Date
2019-12-01
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ISSN
18670342
18670334
18670334
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2-s2.0-85069687819
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Mahidol University
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SCOPUS
Bibliographic Citation
Food and Environmental Virology. Vol.11, No.4 (2019), 374-382
Suggested Citation
Thamapan Tunyakittaveeward, Kitwadee Rupprom, Kannika Pombubpa, Nopporn Howteerakul, Leera Kittigul Norovirus Monitoring in Oysters Using Two Different Extraction Methods. Food and Environmental Virology. Vol.11, No.4 (2019), 374-382. doi:10.1007/s12560-019-09396-y Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/49695
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Norovirus Monitoring in Oysters Using Two Different Extraction Methods
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Abstract
© 2019, Springer Science+Business Media, LLC, part of Springer Nature. Detection of noroviruses in bivalve shellfish is difficult because of the low concentration of norovirus and the presence of reverse transcription (RT)-PCR inhibitors. This study aimed to assess the presence of noroviruses in oysters extracted using a proteinase K extraction (ISO 15216 method) and an adsorption–elution method. Seventy oyster samples were extracted using the two extraction methods and evaluated using RT-nested PCR. The results showed norovirus detection rates at an equal frequency of 28.6%, of which a total of 48 (68.6%) samples had corresponding positive or negative results, while there were 22 (31.4%) samples with discrepant results. Norovirus genogroup (G)I, GII, and mixed GI and GII were detected in 20%, 4.3%, and 4.3% of samples, respectively, by the proteinase K extraction method, which comprised of GI.2, GI.5b, GI.6b, GII.4, and GII.17 genotypes. With the adsorption–elution method noroviruses were detected in 17.1%, 8.6%, and 2.9% of samples, respectively, which comprised of GI.2, GII.2, GII.4, and GII.17 genotypes. All norovirus-positive oyster samples were further estimated for genome copy number using RT-quantitative PCR. The oyster samples processed using the adsorption–elution method contained norovirus GI of 3.36 × 101–1.06 × 105 RNA copies/g of digestive tissues and GII of 1.29 × 103–1.62 × 104 RNA copies/g. Only GII (2.20 × 101 and 7.83 × 101 RNA copies/g) could be quantified in samples prepared using the proteinase K extraction method. The results demonstrate the different performance of the two sample-processing methods, and suggest the use of either extraction method in combination with RT-nested PCR for molecular surveillance of norovirus genotypes in oysters.