Abstract
Foodbome illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25 g sample of whole oyster and lettuce in 175 mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at 4°C was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 pm syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.
| Original language | English |
|---|---|
| Pages (from-to) | 71-76 |
| Number of pages | 6 |
| Journal | Korean Journal of Food Science and Technology |
| Volume | 39 |
| Issue number | 1 |
| Publication status | Published - 2007 Dec 1 |
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Keywords
- Feline calicivirus
- Lettuce
- Norovirus
- Oyster
- Rapid detection
- RT-PCR
ASJC Scopus subject areas
- Food Science
- Biotechnology
- Applied Microbiology and Biotechnology
Cite this
Development of protocol for the effective detection of feline calicivirus as norovirus surrogate in oyster and lettuce. / Lee, Soo Yeon; Jang, Keum Il; Woo, Gun-Jo; Kwak, Hyo Sun; Kim, Kwang Yup.
In: Korean Journal of Food Science and Technology, Vol. 39, No. 1, 01.12.2007, p. 71-76.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of protocol for the effective detection of feline calicivirus as norovirus surrogate in oyster and lettuce
AU - Lee, Soo Yeon
AU - Jang, Keum Il
AU - Woo, Gun-Jo
AU - Kwak, Hyo Sun
AU - Kim, Kwang Yup
PY - 2007/12/1
Y1 - 2007/12/1
N2 - Foodbome illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25 g sample of whole oyster and lettuce in 175 mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at 4°C was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 pm syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.
AB - Foodbome illness caused by Noroviruses (NVs) is increasing rapidly in Korea. This study developed an effective detection protocol for NVs found in contaminated oysters and lettuce through an investigation using the major steps of virus particle separation, concentration and RT-PCR. As a surrogate model for NVs, the cultivable feline calicivirus (FCV) that belongs to the same Caliciviridae family was used. Instead of using a time-consuming ultracentrifugation method, efficient methods based on solvent extraction and PEG precipitation procedure were applied. Direct homogenization of a 25 g sample of whole oyster and lettuce in 175 mL PBS provided the simplicity that would be needed in the actual field of food product examination. The overnight PEG precipitation step at 4°C was reduced to 3 h by placing the reaction tube in ice and by adjusting the PEG concentrations. The application of the use of chloroform and 0.2 pm syringe filtration together showed a better detection efficiency than the use of chloroform alone in removing PCR inhibitors for both oyster and lettuce samples. Also, dilution of the extracted RNA solution before PCR provided increased sensitivity. The improved detection protocol developed in this study could be efficiently applied to detect FCV and most likely NVs from oysters and lettuce.
KW - Feline calicivirus
KW - Lettuce
KW - Norovirus
KW - Oyster
KW - Rapid detection
KW - RT-PCR
UR - http://www.scopus.com/inward/record.url?scp=49649117408&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=49649117408&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:49649117408
VL - 39
SP - 71
EP - 76
JO - Korean Journal of Food Science and Technology
JF - Korean Journal of Food Science and Technology
SN - 0367-6293
IS - 1
ER -