鹅细小病毒于番鸭成纤维细胞上的适应性研究

朱春华; 黄瑜; 傅光华; 万春和; 程龙飞; 傅秋玲

doi:10.19303/j.issn.1008-0384.2014.09.001

鹅细小病毒于番鸭成纤维细胞上的适应性研究

doi: 10.19303/j.issn.1008-0384.2014.09.001

福建省农业科学院畜牧兽医研究所,福建福州,350013

基金项目:

国家公益性行业农业科研专项 (201003012)

详细信息

作者简介:
朱春华 (1980-), 女, 硕士, 助理研究员, 主要从事动物病毒学研究;黄瑜 (1966-), 男, 博士, 研究员, 主要从事动物传染病研究 (E-mail:huangyu_815@163.com)

中图分类号: S852.659.2
计量
- 文章访问数: 93
- HTML全文浏览量: 23
- PDF下载量: 0
- 被引次数: 0
出版历程
- 收稿日期: 2014-04-02
- 刊出日期: 2014-09-18

Adaptation of Goose Parvovirus to Muscovy Duck Embryo Fibroblast

Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences

摘要

摘要: 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。
- 鹅细小病毒 /
- 番鸭成纤维细胞 /
- 增殖 /
- 适应性
Abstract: 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 genomic 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.
- GPV /
- MDEF /
- propagation /
- adaptation

HTML全文

参考文献(18)

[1]	SAIF Y M.禽病学[M].北京:中国农业出版社, 2012:459-467.
[2]	方定一.“小鹅瘟”的介绍[J].中国畜牧兽医杂志, 1962, 9 (8) :19-20.
[3]	胡桂学, 高光, 邹啸环, 等.小鹅瘟病毒分离与初步鉴定[J].经济动物学报, 2002, 6 (2) :41-43.
[4]	KISARY J, DERZSY D.Viral disease of Goslings.IV.Characterization of the causal agent in tissue culture system[J].Acta Vet Acad Sci Hung, 1974, 24 (3) :287-292.
[5]	DERZSY D.A viral disease of goslings.I.Epidemiological, clinical, pathological and aetiological studies[J].Acta Vet Acad Sci Hung, 1967, 17 (4) :443-448.
[6]	GOUGH R E, SPACKMAN D, COLLINS M S.Isolation and characterisation of a parvovirus from goslings[J].Vet Rec, 1981, 108 (18) :399-400.
[7]	POONIA B, DUNN P A, Lu H, et al.Isolation and molecular characterization of a new Muscovy duck parvovirus from Muscovy ducks in the USA[J].Avian Pathol, 2006, 35 (6) :435-441.
[8]	殷震, 刘景华.动物病毒学[M].北京:科学出版社, 1997:581.
[9]	SUVOROV A V.Cytopathic changes produced in goose fibroblast cultures by goose parvovirus[J].Bull Vses Inst Eksp Vet, 1982, 48:16-18.
[10]	王燕玲.番鸭小鹅瘟病免疫荧光诊断方法的建立及应用[D].福州:福建农林大学, 2011.
[11]	朱小丽, 陈少莺, 程晓霞, 等.番鸭小鹅瘟病直接荧光诊断试剂的研制[J].畜牧兽医杂志, 2012, 31 (4) :1-3.
[12]	陈少莺, 胡奇林, 程晓霞, 等.鹅细小病毒弱毒株选育的研究[J].中国预防兽医学报, 2002, 24 (4) :47-49.
[13]	王春媛.鹅细小病毒98E鸭胚致弱株的培育及生物学特性研究[D].哈尔滨:东北农业大学, 2012.
[14]	万春和, 朱海侠, 黄瑜, 等.一株鹅细小病毒全基因特征分析[J].中国动物传染病学报, 2011, 19 (4) :19-24.
[15]	ALEXANDROV M, ALEXANDROVA R, ALEXANDROV I, et al.Fluorescent and electron-microscopy immunoassays employing polyclonal and monoclonal antibodies for detection of goose parvovirus infection[J].J Virol Methods, 1999, 79 (1) :21-32.
[16]	TAKEHARA K, HYAKUTAKE K, IMAMURA T, et al.Isolation, identification, and plaque titration of parvovirus from Muscovy ducks in Japan[J].Avian Dis, 1994, 38 (4) :810-815.
[17]	尹青.蛋用型鸡禽白血病的病毒感染动态及病理学研究[D].泰安:山东农业大学, 2011.
[18]	ZADORI Z, STEFANCSIK R, RAUCH T, et al.Analysis of the complete nucleotide sequences of goose and muscovy duck parvoviruses indicates common ancestral origin with adenoassociated virus 2[J].Virology, 1995, 212 (2) :562-573.