Preliminary Study of RT-LAMP Method for Detection of Ibaraki Virus

To detect Ibaraki virus (IBAV) sensitively and quickly, a rapid detection technique of reverse transcriptase isothermal nucleic acid amplification (RT-LAMP) for the virus was established, through the main steps as follows: the design of 4 specific primers basing on the 6 conserved regions of IBAV VP3 protein gene, the optimization of reaction time and temperature, sensitivity test and specific test. The results showed that when the annealing temperature was 62 °C and the reaction time was 60 min, the electrophoresis band was the most obvious. The method has good specificity for IBAV without cross reaction to bovine epidemic heat virus (BEFV) and bovine viral diarrhea virus (BVDV). At the same time, the sensitivity is 100 times higher than that of RT-PCR, and the concentration of IBAV gene can be detected to 10 copies/μL. The method can complete the amplification reactiononly with a water bath within 1 hour, simply and efficiently. It provides a new detection method for the preliminary screening and detection of IBAV at the base level.