Detection of Venezuelan equine encephalitis virus by reverse transcription loop-mediated isothermal amplification

VernacularTitle:建立快速检测委内瑞拉马脑炎的RT-LAMP技术
Author: Yang CAO ¹ ; Huan LI ; Longfei FENG ; Luping CHEN ; Boran HU ; Jing YUAN
Author Information

1. 扬州大学旅游烹饪学院食品科学与工程学院 225009
Keywords: Reverse transcription loop-mediated isothermal amplification; Venezuelan equine encephalitis virus; Rapid diagnosis; nsp1 gene
From: Chinese Journal of Experimental and Clinical Virology 2020;34(1):78-82
CountryChina
Language:Chinese
Abstract: Objective The study aimed to establish a rapid method for detection of Venezuelan equine encephalitis virus (VEEV) by reverse transcription loop-mediated isothermal amplification (RT-LAMP).Methods Firstly,5 sets of specific primers were designed for the nsp1 gene sequence of VEEV.We screened for the best primer from the 5 sets of primers.After the best primer was obtained,the temperature gradient experiment was carried out.Then,11 viruses were selected for specific test of VEEV.Finally,RT-LAMP and RT-qPCR were compared for sensitivity.Results The optimum temperature for VEEV detection by RT-LAMP was 65 ℃.Amplification result of 10 kinds of virus were negative and VEEV were positive.The result showed that the method has good specificity.It was found that the of RT-LAMP was 100 times higher than that of RT-qPCR.Conclusions The detection method of RT-LAMP is not only simple,rapid aud efficient,but also has high specificity and high sensitivity.