TỐI ƯU HÓA PHẢN ỨNG RT-LAMP PHÁT HIỆN NHANH NOROVIRUS TRONG THỰC PHẨM

Abstract

Norovirus, a virus of the Caliciviridae family, is the leading agent of food borne viral gastroenteritis worldwide. The most common foods contaminated by Norovirus are bivalve molluscan and raw vegetables. Among methods to detect Norovirus, RT-LAMP method is one of the most efficient low-costed isothermal amplifying techniques that provide rapid and exact specification. However the LAMP primer set does not always have sufficient specificity to detect all the various type of Norovirus. Therefore this study was conducted to optimize the LAMP primer set and RT-LAMP method. With the purpose of accomplishing this goal, the food samples which were collected from some markets in Hanoi were processed, followed by extraction and purification of viral RNA, then the RT-LAMP reaction was tested to select optimal conditions (concentration of reverse transcriptase enzyme: 2U / reaction; annealing temperature: 63°C; concentration of betaine for GI: 1M, for GII: 0.8M; incubation time: 60 minutes) with detective sensitivity 10 copies/reaction and specificity of 100% for tested strains.