摘要
采集南京某宠物医院处置的疑似患猫杯状病毒病的家猫的鼻咽拭子,将其处理液接种于CRFK细胞,进行连续传代培养,出现特征性细胞病变。通过对该分离株进行形态学观察、PCR鉴定、间接免疫荧光试验,证明该分离株为猫杯状病毒(FCV),遂将其命名为FB-NJ-13。用RT-PCR分段扩增病毒基因组并将序列提交至GenBank(登录号:KM111557)。序列分析显示,该毒株与国内外参考毒株核苷酸序列的同源性在76.2%~79.6%之间。同时研究了该病毒对温度、酸碱、有机溶剂、干燥和紫外线照射等理化因素的抵抗力。结果显示,50℃30min或70℃5min即可灭活该病毒;该病毒在pH值为3的酸性环境中和pH值为10的碱性环境中不稳定;它对乙醚和氯仿不敏感;紫外线照射2.0h以上,可将该病毒有效灭活。上述研究结果为揭示我国FCV的分子生物学特征和遗传进化规律提供了重要的科学依据。
Nasopharyngeal swabs were collected from an infected cat,which was suspected with feline calicivirus(FCV)infection and was treated in a pet hospital in Nanjing,China,and then were inoculated into CRFK cells.The cultured cells were passaged continually.Finally,the cell lines had a cytopathic effect.Through morphological observation,PCR identification,and indirect immunofluorescent assay,the isolate was proved to be feline calicivirus,then temporarily named as FB-NJ-13.The virus complete genome was amplified by RT-PCR and was submitted to GenBank(Accession number:KM111557).Sequence analysis showed that the nucleotide sequence homology between the isolate and reference strains was from 76.2%to 79.6%.Studies on the detailed physical and chemical properties(including temperature,pH,organic solvent,dry and ultraviolet irradiation)of the isolate were carried out.In result,the isolate was inactivated at50℃for 30 min or at 70℃for 5min.It was unstable in the acidic solution of pH3 and in the alkaline solution of pH10,and was insensitive to ether and chloroform.It was inactivated while the ultraviolet irradiation was done for more than 2.0h.In conclusion,the isolation and identification of the feline calicivirus strain has an important significance on further study of FCV molecular biology and genetic evolution.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2015年第4期385-389,共5页
Chinese Veterinary Science
基金
中央级公益性科研院所基本科研业务费项目(0302014012)
关键词
猫杯状病毒
分离鉴定
理化因素
全基因组
feline calicivirus
isolation and identification
physical and chemical factors
complete genome
