Abstract:

Reverse transcription polymerase chain reaction (RT-PCR), RT-seminested PCR and reverse transcription loopmediated isothermal amplification (RT-LAMP) assays were compared to detect norovirus in oysters. All the three assays targeted the highly conserved capsid N-terminal/shell (N/S) domain of norovirus with specific primers. Although one-step RT-PCR presented more apparent results than two-step approach, it still had the limitation in eliminating inhibitors contained in food matrix, and both RT-seminested PCR and RT-LAMP showed significantly higher specificity and sensitivity than RT-PCR. However, the operation of RT-seminested PCR is tedious and time-consuming. In comparison to other two approaches, the amplification program of RT-LAMP is simple, and the reaction time is short. Without special apparatus needed, RT-LAMP can be performed under isothermal condition and the results may be visually ascertained by a simple color reaction using SYBR GreenⅠdye. Therefore, RT-LAMP is of great potential to become a valuable method for rapid detection of norovirus in oysters.

Key words: norovirus, oyster, detection, RT-PCR, RT-seminested PCR, RT-LAMP