不同毒力水貂阿留申病毒在猫肾细胞中的增殖特性及凋亡特性的比较研究

发布时间：2018-04-16 21:27

本文选题：水貂阿留申病毒(AMDV) + 猫肾细胞(CPFK)　；参考：《中国畜牧兽医》2017年09期

【摘要】：试验旨在对不同毒力水貂阿留申病毒(Aleutian mink disease virus,AMDV)在猫肾细胞(feline kidney cell,CRFK)中的增殖规律及其诱导细胞凋亡情况进行比较研究。将标准毒株AMDV-G及分离到的野毒株AMDVDL124、AMDV-DL125、AMDV-QD2、AMDV-QD3、AMDV-ZJ3接种CRFK细胞,应用间接免疫荧光、实时荧光定量PCR、TCID50测定技术研究病毒在细胞中的复制及表达情况,同时检测病毒诱导的细胞凋亡情况。间接免疫荧光结果显示,5株野毒株荧光着色趋势差异不大,均在感染后12h出现荧光,随感染时间延长荧光增多,AMDV-G荧光出现时间比野毒株晚,但病毒感染后72h几乎所有细胞均出现荧光;实时荧光定量PCR结果显示,基因组复制趋势大致相同,AMDV-DL125感染后3h复制开始,AMDV-G感染后24h复制才开始并呈快速增长趋势,但感染后72h均达到峰值。TCID_(50)检测结果表明,0~12h为病毒感染潜伏期,AMDV-G感染后60h达到峰值,野毒株均在感染后72h达到峰值,但是6株病毒均能在感染后48~72h维持较高的感染滴度,其后随细胞崩解而降低。SPSS 23.0统计软件分析凋亡检测结果显示,与对照组相比,野毒株感染细胞后2~12h诱导细胞凋亡差异显著(P0.05),AMDV-G诱导细胞凋亡差异明显低于野毒株,但是诱导细胞凋亡时间较野毒株长,在感染后24h仍对细胞凋亡有较明显的诱导作用,但是各病毒诱导的细胞凋亡主要集中在2~12h。该结果为AMDV的培养、鉴定及致病机理研究提供一定参考。
[Abstract]:The aim of this study was to compare the proliferation and apoptosis of Aleutian mink disease virus (Aleutian mink disease virus) in feline kidney cell line (CRFK) of different virulent mink cells.AMDV-QD2AMDV-QD3AMDV-ZJ3 was inoculated with standard AMDV-G and AMDV-DL124AMDV-DL125AMDV-QD3AMDV-ZJ3. The replication and expression of AMDV-QD2AMDV-QD3AMDV-ZJ3 in CRFK cells were studied by real-time fluorescence quantitative PCR TCID50 assay, and the cell apoptosis induced by AMDV-QD2AMDV-QD2AMDV-QD2AMDV-QD3AMDV-ZJ3 was detected.The results of indirect immunofluorescence showed that there was no significant difference in fluorescent staining of 5 wild strains, all of them appeared fluorescence 12 hours after infection, and the fluorescence time of AMDV-G was later than that of wild strain with the increase of infection time.The results of real-time fluorescence quantitative PCR showed that the trend of genome replication was approximately the same. The replication of AMDV-DL125 began at 3h after infection, and the replication of AMDV-G began only 24 hours after infection and showed a rapid increasing trend, the results of real-time fluorescence quantitative PCR showed that the genome replication was similar to that of AMDV-DL125 infection at 3 hours after infection.However, the peak value was reached at 72 h after infection. The results showed that the peak value of AMDV-G was reached at 12 h after infection and that of wild strain reached the peak at 72 h after infection. However, all the 6 strains could maintain high titer of infection at 48 h 72 h after infection, and AMDV-G reached the peak value at 60 h after infection, and all the wild strains reached the peak at 72 h after infection, but all the 6 strains were able to maintain a high titer of infection at 48 h after infection.The results of apoptosis analysis with SPSS23.0 statistical software showed that the difference of apoptosis induced by AMDV-G was significantly lower than that of the control group at 212h after infection with AMDV-G.But the time of inducing cell apoptosis was longer than that of wild virus strain, and still had obvious effect on cell apoptosis at 24 h after infection, but the apoptosis induced by each virus was mainly concentrated in 2o 12h.The results provide some references for the culture, identification and pathogenesis of AMDV.
【作者单位】：吉林农业大学动物科学与技术学院;吉林农业大学研究生院;
【基金】：产业创新战略联盟项目(20140309018YY) 吉林省科技厅科技成果转化促进计划(20140412009XH)
【分类号】：S852.65

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