抗低温消毒剂“抗冻消-30”对非洲猪瘟病毒的杀灭效果

Inactivation of African swine fever virus by anti-cryogenic disinfectant"Kangdongxiao-30"

为评估抗低温消毒剂"抗冻消-30"对非洲猪瘟病毒(ASFV)的杀灭效果,本研究通过观察不同稀释度的"抗冻消-30"对细胞形态的影响,并计算相应细胞的活性确定该消毒剂对细胞的安全浓度。结果显示,"抗冻消-30"对细胞的安全浓度为≤1 g/L,细胞存活率高于95%,且形态正常。将不同浓度的ASFV报告病毒(rASFV-HLJ/2018-EGFP株)与不同浓度"抗冻消-30"在不同温度下作用30 min后,分别感染HEK293T细胞,通过倒置荧光显微镜观察,评估在不同温度下"抗冻消-30"对ASFV报告病毒的杀灭效果;将不同浓度的ASFV亲本病毒(HLJ/2018株)与不同浓度"抗冻消-30"在-30℃作用30 min后,分别感染猪肺泡巨噬细胞(PAMs),一定时间后加入猪红细胞,利用倒置显微镜观察红细胞吸附现象,评估-30℃条件下,"抗冻消-30"对ASFV亲本病毒的杀灭效果。结果显示,常温和4℃时,400 g/L、160 g/L、80 g/L、40 g/L、8 g/L的"抗冻消-30"可完全杀灭106TCID50及以下浓度、105TCID50及以下浓度、1104TCID50及以下浓度、104TCID50及以下浓度、103TCID50及以下浓度的ASFV报告病毒;-20℃时,上述浓度的"抗冻消-30"可完全杀灭105TCID50及以下浓度、1104TCID50及以下浓度、103TCID50及以下浓度、103TCID50及以下浓度、102TCID50 ASFV报告病毒;-30℃和-40℃时,上述浓度的"抗冻消-30"可完全杀灭1104TCID50及以下浓度、103TCID50及以下浓度、103TCID50及以下浓度、102TCID50及以下浓度、102TCID50 ASFV报告病毒。-30℃时,400 g/L、160 g/L、80 g/L、40 g/L的"抗冻消-30"可完全杀灭1104TCID50及以下浓度、103TCID50及以下浓度、103TCID50及以下浓度、102TCID50及以下浓度的ASFV亲本病毒。本研究在细胞水平上系统评价了该消毒剂对ASFV报告病毒和亲本病毒的杀灭情况,表明"抗冻消-30"在常温和低温条件下对ASFV均具有良好的杀灭效果,可以作为防控ASF的有效消毒剂。

In order tosystematically evaluate the inactivation effect of"Kangdongxiao-30"on African swine fever virus(ASFV),in this study,the safe concentration of"Kangdongxiao-30"was determined by observing the effect of disinfectant on cell morphology of monolayer cells and calculating the cell activity.The results showed that the safe concentration of"Kangdongxiao-30"for the cell was≤1 g/L,and the cell viability was higher than 95%and the cell morphology was not affected during using the concentration.The inactivation effect of"Kangdongxiao-30"on reporter ASFV(r ASFV-HLJ/2018-EGFP strain)at different temperatures was evaluated,the reporter ASFV was treated with different concentrations of"Kangdongxiao-30"at different temperatures for 30 min,and then inoculate HEK293 T cells,these cells were observed by inverted fluorescence microscope.Meanwhile,the inactivation effect of"Kangdongxiao-30"on parental ASFV(HLJ/2018 strain)at-30℃was also evaluated,the parental ASFV was treated with different concentrations of"Kangdongxiao-30"for 30 min,and then inoculate PAMs,the hemadsorption phenomenon of these cells was observed by inverted fluorescence microscope after porcine red blood cells were added.The results showed that the reporter ASFV of≤106TCID50,≤105TCID50,≤104TCID50,≤104TCID50,≤103TCID50 were completely inactivated by 400 g/L,160 g/L,80 g/L,40 g/L,8 g/L"Kangdongxiao-30"at room temperature and 4℃,respectively.The reporter ASFV of≤105TCID50,≤104TCID50,≤103TCID50,≤103TCID50,≤102TCID50 were completely inactivated by the above concentrations of"Kangdongxiao-30"at-20℃,respectively.The reporter ASFV of≤104TCID50,≤103TCID50,≤103TCID50,≤102TCID50,≤102TCID50 were completely inactivated by the above concentrations of"Kangdongxiao-30"at-30℃and-40℃,respectively.Besides,the parental ASFV of≤104TCID50,≤103TCID50,≤103TCID50,≤102TCID50 were completely inactivated by 400 g/L,160 g/L,80 g/L,40 g/L"Kangdongxiao-30"at-30℃,respectively.The study proved that"Kangdongxiao-30"had a good inactivation effect on ASFV at room temperature and under cryogenic condition,and it could be used as an effective disinfectant to inactivate ASFV.