Influence of PCR Inhibitor Removal on Detection of Norovirus GⅡ in Frozen Strawberry by
Real-Time Fluorescent RT-PCR
XU Wenfei, FANG Baohai, LIN Chao, WANG Gongpu, WANG Qun, ZHENG Xiaolong

doi:10.7506/spkx1002-6630-201704048

Abstract

Abstract: Objective: Comparative experiments were designed to address the influence of the presence of PCR inhibitor on the results of detection of norovirus GⅡ in frozen strawberry by real-time fluorescent RT-PCR (RT-qPCR), aiming to improve the detection sensitivity and accuracy. Methods: After pretreatment of fecal suspensions containing norovirus GⅡ and known negative strawberry samples, the virus was enriched for the extraction and purification of RNA and a control group and an experimental group were established. After removal of the PCR inhibitor, the samples were determined by RTqPCR. Results: The effect of the inhibitor tended to increase obviously with decreasing concentration gradient of virus, thus giving false negative results which were two orders of magnitude higher than the lowest level detected without the presence of the inhibitor. Better detection results were obtained by removal of the inhibitor compared with the control untreated group, as the virus concentration gradient decreased. The stable detection limit (LOD) for the experimental group without PCR inhibitor was 10-6 (Ct value < 40), and the minimum LOD were 10-7. Conclusion: The detection of norovirus in frozen strawberries by fluorescence quantitative RT-PCR could be improved in terms of sensitivity by removal of the inhibitor. In addition, the improved method was quick, easy to operate, economic and practical.

Key words: frozen strawberry, norovirus G Ⅱ, RT-PCR, inhibitors

CLC Number:

X954

XU Wenfei, FANG Baohai, LIN Chao, WANG Gongpu, WANG Qun, ZHENG Xiaolong, LIANG Chengzhu, YUE Zhiqin. Influence of PCR Inhibitor Removal on Detection of Norovirus GⅡ in Frozen Strawberry by Real-Time Fluorescent RT-PCR XU Wenfei, FANG Baohai, LIN Chao, WANG Gongpu, WANG Qun, ZHENG Xiaolong[J]. FOOD SCIENCE, 2017, 38(4): 296-300.

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Influence of PCR Inhibitor Removal on Detection of Norovirus GⅡ in Frozen Strawberry by Real-Time Fluorescent RT-PCR XU Wenfei, FANG Baohai, LIN Chao, WANG Gongpu, WANG Qun, ZHENG Xiaolong

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