浒苔多糖体外抗犬冠状病毒临床分离株的作用研究

Study on Anti-CCoV Clinical Isolated Strain by Enteromorpha Polysaccharide in Vitro

利用A72细胞(犬纤维肉瘤细胞系)从山东某宠物医院临床上表现为急性胃肠炎症状的幼犬腹泻粪便中分离到1株病毒,通过间接免疫荧光(IFA)和RT-PCR进行鉴定,确定为犬冠状病毒,命名为CCoV-SW1。为探讨浒苔多糖(EPS)体外抗CCoV-SW1毒株的作用效果,以A72细胞为细胞模型,通过直接杀灭、抑制复制及阻断吸附3个方面进行研究。结果表明,EPS对A72细胞最大安全浓度为62.5mg/L;EPS对CCoV感染A72细胞抑制率与EPS质量浓度有一定量效关系;EPS对CCoV具有直接杀灭和阻断吸附的作用,并且与EPS浓度具有量效关系;EPS抑制CCoV复制效果不明显,但仍有一定效果,当EPS浓度为62.5mg/L时,细胞存活率为37.2%。EPS在体外对CCoV具有直接杀灭和阻断吸附的作用,为CCoV的预防和治疗提供依据。

An CCoV strain was isolated by using A72 cells from the taeces ot a pup with acute gastroenterltis in a pet hospital of Shandong.Through the identification by IFA and RT-PCR, the results demonstrated that the iaolated virus is CCoV,named CCoV-SWI.In order to explore the antiviral effects of Enterornorpha polysaccharide（EPS）, the following experiments,including the directly viruscidal assay, virus inhibition replication assay and virus blocking absorbtion assay with A72 cell model were used.The results indicated that the maximum safety concentration of EPS to A72 cells was 62.5 mg/L;the inhibition ratio to CCoV infected A72 cells of EPS had a quatitative relationship with the EPS concentration; the EPS had a strong anti-CCoV ability in vitro by directly virucidal effect and blocking absorbtion of the virus to cells,it had a quatitative relationship with the EPS concentration;the effects of EPS to inhibit replication of CCoV was not obvious, hut still had some effects,when mass concentration of EPS was 62.5 mg/L, the inhibition ratio of cells was 37.2%. This study indicated that the EPS has a strong anti-CCoV ability in vitro by directly virucidal effect and blocking ahsorbtion of the virus to cells,it can provide important scientific data for preventing and treating CCoV.