Study on Anti-Novel duck reovirus Effect of Chlorogenic Acid In vitro
PAN Li1,2*, MA Xiu-Li1*, HUANG Zhong-Li1, LI Gui-Ming1, TANG Liang1,2, YU Ke-Xiang1**, LIN Shu-Qian1**
1 Institute of Poultry, Shandong Academy of Agricultural Sciences, Jinan 250023, China; 2 College of Animal Science and Technology, Shandong Agricultural University, Tai'an 271018, China
Abstract：Novel duck reovirus (NDRV) disease is a serious infectious disease for poultry in China. Chlorogenic acid (CHA) is a main component of many traditional Chinese medicines, and plays an important role in antiviral research. Exploring the antiviral effect of CHA on NDRV which might provide a basis for the clinical treatment of NDRV to some extent. Firstly, based on S3 gene sequence (GenBank No. KJ879932) of NDRV, the method of real-time fluorescent quantitative PCR (qPCR) was established. Secondly, maximum safe concentration of CHA on baby hamster (Mesocricetus auratus) kidney cell line BHK-21 cells was determined by CCK-8 kit. Finally, the anti-virus test was performed. The experiment was divided into 2 groups. GroupⅠ: NDRV were inoculated in BHK-21 cells before different concentrations of CHA were added. Then the freeze-thaw solution of cells was collected periodically. GroupⅡ: The method of virus adsorption was the same as that of groupⅠ. Before treatment, the virus were proliferated on BHK-21 cells for 10 h, and the viral load after 10 h proliferation was quantitatively detected as the base value. After 10 h of virus proliferation, different concentrations of CHA were added, and the follow-up procedure was the same as that of Group Ⅰ. Negative and positive control were set in both groups. And the remaining experiment procedures were the same as those in groupⅠ. The cytopathic effects of each group were observed regularly, and the collected samples were quantitatively detected by qPCR. The results showed the lowest detectable viral nucleic acid of the method was 92.5 copies/μL, and the maximum safe concentration of CHA on BHK-21 cells was 128 μg/mL. The effect of anti-NDRV of CHA indicated that different concentrations of CHA could inhibit the proliferation of NDRV in vitro, and this inhibitory effect had an obvious dose-effect relationship. This study could provide basic data for the study of NDRV mechanism and its practical application.
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