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http://dx.doi.org/10.13103/JFHS.2014.29.1.031

Quantitative Analysis of Feline Calicivirus Inactivation using Real-time RT-PCR  

Jeong, Hye Mi (Dept. of Food Science and Biotechnology, Chungbuk National University)
Kim, Kwang Yup (Dept. of Food Science and Biotechnology, Chungbuk National University)
Publication Information
Journal of Food Hygiene and Safety / v.29, no.1, 2014 , pp. 31-39 More about this Journal
Abstract
Norovirus causes acute gastroenteritis in all age groups and its food poisoning outbreaks are rapidly increasing in Korea. Reverse transcription-polymerase chain reaction (RT-PCR) is most widely used for the rapid detection of foodborne viruses due to high sensitivity. However, the false positive results of RT-PCR obtained against already inactivated viruses could be a serious drawbacks in food safety area. In this study, we investigated a method to yield true positive RT-PCR results only with alive viruses. To decompose the RNA genes from dead viruses, the enzymatic treatments composed of proteinse K and Ribonuclease A were applied to the sanitized and inactivated virus particles. Another aim of this study was to quantify the efficiencies of several major sanitizing treatments using real-time RT-PCR. Feline calicivirus (FCV) that belongs to the same Caliciviridae family with norovirus was used as a surrogate model for norovirus. The initial level of virus in control suspension was approximately $10^4$ PFU/mL. Most of inactivated viruses treated with the enzymatic treatment for 30 min at $37^{\circ}C$ were not detected in RT-PCR, Quantification results to verify the inactivation efficiencies of sanitizing treatments using real-time RT-PCR showed no false positive in most cases. We could successfully develope a numerical quantification process for the inactivated viruses after major sanitizing treatments using real-time RT-PCR. The results obtained in this study could provide a novel basis of rapid virus quantification in food safety area.
Keywords
Norovirus; Feline Calicivirus; Inactivation; Enzymatic treatment; Real-time RT-PCR;
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