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Rapid Detection of Norovirus from Fresh Lettuce Using Immunomagnetic Separation and a Quantum Dots Assay

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Abstract:

Current molecular methods that include PCR have been used to detect norovirus in many food samples. However, the protocols require removing PCR inhibitors and incorporate time-consuming concentration steps to separate virus from analyte for rapid and sensitive detection of norovirus. We developed an immunomagnetic separation (IMS) and a quantum dots (QDs) assay to detect norovirus eluted from fresh lettuce with Tris buffer containing 1% beef extract (pH 9.5). IMS facilitated viral precipitation with a 10-min incubation, whereas virus concentration using polyethylene glycol (PEG) requires more than 3 h and an additional high-speed centrifugation step to precipitate virus before reverse transcription PCR (RT-PCR) analysis. The fluorescence intensity of QDs was detected qualitatively on norovirus dilutions of 10–1 to 10–3 in a stool suspension (100 RT-PCR units/ml). The results suggest that a fluorescence assay based on IMS and QDs is valid for detecting norovirus qualitatively according to fluorescent signal intensity within the same virus detection limit produced by IMS–RT-PCR and PEG–RT-PCR.

Document Type: Research Article

DOI: http://dx.doi.org/10.4315/0362-028X.JFP-12-343

Affiliations: 1: Department of Food Science and Technology and Functional Food Research Center, Chonnam National University, Gwangju 500-757, Republic of Korea 2: Korea Basic Science Institute, Gwangju 500-757, Republic of Korea 3: Korea Basic Science Institute, Daejeon 305-806, Republic of Korea 4: Department of Rural and Biosystems Engineering, Chonnam National University, Gwangju 500-757, Republic of Korea 5: School of Mechatronics, Gwangju Institute of Science and Technology, Gwangju 500-757, Republic of Korea 6: Department of Aqualife Medicine, Chonnam National University, Yeosu 550-749, Jeonnam, Republic of Korea 7: Health & Environment Institute of Gwangju, 898 Hwajung-dong, Seo-gu, Gwangju 502-837, Republic of Korea 8: Department of Food Science and Technology and Functional Food Research Center, Chonnam National University, Gwangju 500-757, Republic of Korea. dwkim@jnu.ac.kr

Publication date: April 1, 2013

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