Abstract:

A SYBR GreenⅠ real-time polymerase chain reaction (RT-PCR) method using the primers designed to encompassing the conservative region of vp3/vp1 in the HAV was developed for the detection of HAV in strawberry. The limit of detection of the method was 101 TCID50. The standard curve was Ct = －3.052 lg(TCID50 )+ 34.57, displaying good linearity over a linear range from 101 to 105 TCID50 with 0.9961 correlation coefficient. This method was specific to HAV without any cross-reaction with rotavirus, adenovirus, norovirus or astrovirus. The coefficients of variation (CV) for determining HAV standards with different titers by this method were between 0.85% and 1.71% (n = 5) in intra-assay and between 0.76% and 1.98% (n = 3) in inter-assay. Totally 45 strawberry samples were examined, and 3 of them were found positive. This method is sensitive, specific and reliable, and can be used for the rapid detection of HAV in fruits and vegetables.

Key words: HAV, SYBR Green Ⅰ real-time RT-PCR, detection, strawberry