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Volume 7, Issue 6 (Suppl)
J Biotechnol Biomater, an open access journal
ISSN: 2155-952X
World Biotechnology 2017
December 04-05, 2017
2
nd
World Biotechnology Congress
December 04-05, 2017 | Sao Paulo, Brazil
Estimate infectivity of human
Norovirus
in environmental water samples by in situ capture RT-qPCR
method
Peng Tian
Agricultural Research Service, USA
H
uman
Noroviruses
(HuNoVs) are highly infectious viruses for which water is an important medium of transmission. In this study,
we explored a new
in situ
capture RT-qPCR (ISC-RT-qPCR) methodology to estimate the infectivity of HuNoV in environmental
water samples. This assay was based on capturing encapsidated HuNoV by viral receptors, followed by
in situ
amplification of the
captured viral genomes by RT-qPCR. We demonstrated that the ISC-RT-qPCR did not capture and enable signal amplification of
heat-denatured Tulane Virus (TV) and HuNoVs. We further demonstrated that the sensitivity of ISC-RT-qPCR was equal or better
than that of conventional RT-qPCR procedures for the detection of HuNoV GI and GII. We then utilized the ISC-RT-qPCR to
detect HuNoV in environmental water samples for comparison against that from a conventional RT-qPCR procedure. TV was used
as a process control virus. While complete inhibition of TV genomic signal was observed in 27% of samples tested by RT-qPCR, no
inhibition of TV genomic signal was observed by ISC-RT-qPCR. From 72 samples tested positive for HuNoV GI signal by RT-qPCR,
only 20 (27.8%) of these samples tested positive by ISC-RT-qPCR, suggesting that 72.2% of RT-qPCR-positive samples were unlikely
to be infectious. From 16 samples tested positive for HuNoV GII signal by RT-qPCR, only one of these samples tested positive by ISC-
RT-qPCR. Five samples that had initially tested negative for HuNoV GII signal by RT-qPCR, was tested as positive by ISC-RT-qPCR.
Overall, ISC-RT-qPCR method provided an alternative assay to estimate infectivity of HuNoV in environmental samples.
Biography
Peng Tian has his expertise in human
Norovirus
and food safety. He has developed several methods to concentrate and detect human norovirus from environmental and
food samples. These approaches are available to all stakeholders interested in viral contamination in food.
peng.tian@ars.usda.gov.comPeng Tian, J Biotechnol Biomater 2017, 7:6 (Suppl)
DOI: 10.4172/2155-952X-C1-085