本文選題：水貂阿留申病毒(AMDV) + 貓腎細(xì)胞(CPFK)��； 參考：《中國畜牧獸醫(yī)》2017年09期

【摘要】：試驗旨在對不同毒力水貂阿留申病毒(Aleutian mink disease virus,AMDV)在貓腎細(xì)胞(feline kidney cell,CRFK)中的增殖規(guī)律及其誘導(dǎo)細(xì)胞凋亡情況進(jìn)行比較研究。將標(biāo)準(zhǔn)毒株AMDV-G及分離到的野毒株AMDVDL124、AMDV-DL125、AMDV-QD2、AMDV-QD3、AMDV-ZJ3接種CRFK細(xì)胞,應(yīng)用間接免疫熒光、實時熒光定量PCR、TCID50測定技術(shù)研究病毒在細(xì)胞中的復(fù)制及表達(dá)情況,同時檢測病毒誘導(dǎo)的細(xì)胞凋亡情況。間接免疫熒光結(jié)果顯示,5株野毒株熒光著色趨勢差異不大,均在感染后12h出現(xiàn)熒光,隨感染時間延長熒光增多,AMDV-G熒光出現(xiàn)時間比野毒株晚,但病毒感染后72h幾乎所有細(xì)胞均出現(xiàn)熒光;實時熒光定量PCR結(jié)果顯示,基因組復(fù)制趨勢大致相同,AMDV-DL125感染后3h復(fù)制開始,AMDV-G感染后24h復(fù)制才開始并呈快速增長趨勢,但感染后72h均達(dá)到峰值。TCID_(50)檢測結(jié)果表明,0~12h為病毒感染潛伏期,AMDV-G感染后60h達(dá)到峰值,野毒株均在感染后72h達(dá)到峰值,但是6株病毒均能在感染后48~72h維持較高的感染滴度,其后隨細(xì)胞崩解而降低。SPSS 23.0統(tǒng)計軟件分析凋亡檢測結(jié)果顯示,與對照組相比,野毒株感染細(xì)胞后2~12h誘導(dǎo)細(xì)胞凋亡差異顯著(P0.05),AMDV-G誘導(dǎo)細(xì)胞凋亡差異明顯低于野毒株,但是誘導(dǎo)細(xì)胞凋亡時間較野毒株長,在感染后24h仍對細(xì)胞凋亡有較明顯的誘導(dǎo)作用,但是各病毒誘導(dǎo)的細(xì)胞凋亡主要集中在2~12h。該結(jié)果為AMDV的培養(yǎng)、鑒定及致病機(jī)理研究提供一定參考。
[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.
【作者單位】： 吉林農(nóng)業(yè)大學(xué)動物科學(xué)與技術(shù)學(xué)院;吉林農(nóng)業(yè)大學(xué)研究生院;
【基金】：產(chǎn)業(yè)創(chuàng)新戰(zhàn)略聯(lián)盟項目(20140309018YY) 吉林省科技廳科技成果轉(zhuǎn)化促進(jìn)計劃(20140412009XH)
【分類號】：S852.65

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