鹅细小病毒于番鸭成纤维细胞上的适应性研究

为进一步了解鹅细小病毒（GPV）在番鸭成纤维细胞（MDEF）上的适应性及病毒增殖情况,将GPV在MDEF上连续传12代培养,通过PCR、免疫荧光、流式细胞等方法分析其生物学特性。PCR检测结果显示,扩增到大小为734bp的非结构蛋白基因片段,将10倍梯度稀释的病毒悬液接种细胞培养65h后,10-6倍接种的细胞仍能检测到病毒核酸。免疫荧光检测显示,兔抗GPV阳性血清能对染毒的病灶产生特异性荧光染色。流式细胞试验结果进一步证实,GPV在MDEF上增殖速度比较慢,病毒的感染会造成MDEF的早期凋亡。说明该GPV株感染MDEF后虽未产生细胞病变,但病毒能在MDEF中增殖,为GPV开展细胞依赖性相关研究奠定基础。

Adaptation of Goose Parvovirus to Muscovy Duck Embryo Fibroblast

In order to study the adaptation and propagation of goose parvovirus （GPV） in Muscovy duck embryo fibroblast （MDEF）, cultures through serial passages, and PCR assay, indirect immunofluorescence, flow cytometry were used to study the biological characteristics of GPV. Specified genomie nonstructural protein （NS） gene of GPV with the segment lengths of 734 bp was amplified by PCR, and PCR assays showed that the cell cultured 65 h after inoculate with 10 6 dilution of virus suspensions could be recognized specifically. Immunofluorescent studies showed that the infected MDEF had fluorescence characteristics using purified rabbit polyclonal antibody against GPV. The flow cytometry assay demonstrated that the GPV propagated slowly in MDEF, and GPV infection would induce the apoptosis in early stage. The results indicated the GPV strain adapted to and propagated in MDEF without cytopathic effects （CPE） on the cultures. The study provides a basis for cell dependent correlation research of GPV.