Establishment and Preliminary Application of Droplet Digital RT-PCR Assay for Detection of GⅡ Norovirus
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Abstract:
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.
