Abstract: Objective: Different procures for viral concentration and RNA extraction from fruits and vegetables were proposed taking into account the difference in surface structure between both materials and they were used to develop a sensitive and rapid real-time fluorescent reverse transcription-polymerase chain reaction (RT-PCR) for the quantitative detection of astrovirus in fruits and vegetables using the primers and probes designed according to published sequences. Methods: A standard curve for quantitative detection of astrovirus was constructed by plotting the cycle threshold (Ct value) versus the starting concentration of 10-fold serially diluted cRNA. Then the method for norovirus and hepatitis A virus RNA extraction from fruits and vegetables described in the standard ISO/TS 15216-2:2013 was applied to detect astrovirus. The viral recovery rate, sensitivity and reproducibility of the assay were evaluated by using artificially contaminated cabbage and strawberry samples. Finally, 60 fruit and vegetable samples were tested to demonstrate the feasibility of this method. Results: The amplification efficiency of the real-time fluorescent RT-PCR was 95.9%, with a limit of detection (LOD) of 5.6 copies per reaction. The viral recovery rate of artificially contaminated cabbage samples was 0.94%?9.63%, compared to only 1.03% for the strawberries. Analysis of the artificially contaminated samples containing the same viral levels demonstrated high reproducibility with a coefficient of variation (CV) of less than 2%. Additionally, one strawberry sample collected from a retail market in Beijing was shown to be astrovirus-positive, and the detection rate was 1.67%. Conclusion: The developed method is rapid, efficient and sensitive for quantitative detection of astrovirus in fruits and vegetables, and will be a useful tool for routine screening and risk assessment of foodborne viruses.

Key words: astrovirus, real-time fluorescent RT-PCR, fruits and vegetables, quantitative detection