为了建立快速准确的GⅡ型诺如病毒定量检测方法，本研究将微滴式数字RT-PCR技术应用于GⅡ型诺如病毒检测中，并与实时荧光RT-PCR法进行了比对。微滴式数字RT-PCR反应退火温度的优化实验确定了其最佳退火温度为56 ℃，实时荧光RT-PCR和微滴式数字RT-PCR两种方法的灵敏度实验对比表明微滴式数字RT-PCR法检测GⅡ型诺如病毒的灵敏度为5.40 copies/μL，其灵敏度高于实时荧光RT-PCR法，重复性实验对比表明两种方法在检测中间浓度的GⅡ型诺如病毒时重复性均较好；人工污染西生菜实验中表明微滴式数字RT-PCR法最低检测限为54.00 copies/μL。本研究建立的GⅡ型诺如病毒的微滴式RT-PCR检测方法，灵敏度高，重复性良好，在人工污染西生菜GⅡ型诺如病毒的检测中表现理想，具有良好的应用前景。

The droplet digital RT-PCR assay was applied in the detection of GⅡnorovirus to establish a rapid and accurate method in this study, which was compared with real-time fluorescent RT-PCR. The annealing temperature of droplet digital RT-PCR was optimized and determined to be 56℃. Compared with real-time fluorescent RT-PCR, the sensitivity of droplet digital RT-PCR was determined to be 5.40 copies/μL which was higher than that of real-time fluorescent RT-PCR, and the repeatability of real-time fluorescent RT-PCR and droplet digital RT-PCR were good determined by the comparison test. The droplet digital RT-PCR was applied to detect the artificially contaminated Romaine Lettuce with the sensitivity wof 54.00 copies/μL. Droplet digital RT-PCR assay established for detection of GⅡnorovirus had a high sensitivity and repeatability, and had an outstanding performance in the detection of the artificially contaminated Romaine Lettuce, which would own a good application prospects.

广州市科信委项目（2014J4500027）