Establishment of RT-LAMP detection method for Bluetongue virus

摘要

In order to establish the RT-LAMP detectionmethod of bluetongue virus (BTV), 4 specific LAMP primers were designed and synthesized by using the conservative sequence of the BTV VP7 genome published by Gen Bank. Through the synthesis of BTV artificial genes, the optimization of the reaction conditions of LAMP, the LAMP sensitivity experiment of BTV and the specific experiments, a preliminary method of BTV LAMP detection was established. The results showed that the BTV gene was successfully synthesized and the LAMP reaction conditions were optimized preliminarily, and the detection limit of the developed BTV LAMP method system could reach 43.7 copies/μL of target gene fragment, which could be detected with good sensitivity and specificity. In the experiment, the BTV virus could be amplified with a good specific amplification at 62°C and 60min, and the results were negative after the amplification of DNA of Escherichia coli and Escherichia coli under the system constructed by the experiment.