检测鹅细小病毒（GPV）强毒PCR方法的建立及人工感染GPV强毒在雏鹅体内分布规律的研究
|
摘要
根据国际基因库(GeneBank)1995年发布的鹅细小病毒(GPV)B株的全基因组序列，针对GPV的保守基因片段VP3设计一对引物，建立了一种用于检测GPV的特异的PCR方法，该法能够特异地扩增出预期大小约441bp片段。特异性实验证明，该法对鸭瘟病毒、鹅致病性大肠杆菌O_8和O_1、雏鹅新型病毒性肠炎病毒(NGVEV)呈阴性反应，仅能从鹅细小病毒(GPV)和番鸭细小病毒(MDPV)中扩增出特异性片段。灵敏性实验证明该方法的检测GPV核酸的灵敏度达到0.47pg。
运用所建立的特异的PCR方法，对人工感染鹅细小病毒后病毒在雏鹅体内的分布规律进行了研究，结果表明：病毒在雏鹅体内的分布规律因病毒感染的途径的不同而存在差异。经口感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时能从心脏、肝脏中检出病毒DNA、感染后24小时能从胸腺、胰腺、十二指肠、小肠、法氏囊、心脏、肝脏、脾脏、肾脏、血液中检出病毒DNA，感染后48小时后能够从直肠、骨髓、肺脏中检出病毒DNA，感染后72小时能从小脑、延髓、盲肠、肌肉、粪便中检出病毒DNA，感染后312小时后仍能从十二指肠、空肠、直肠、心脏、肝脏、肾脏、肌肉、粪便中检出病毒DNA。经皮下注射感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时能够从心脏，肝脏中检测出病毒DNA，感染后24小时能够从延髓、胸腺、胰腺、十二指肠、空肠、盲肠、法氏囊、心脏、肝脏、脾脏、肺脏、肾脏、血液、小脑、直肠、肌肉、粪便中检出病毒DNA，感染后48小时后能从骨髓中检测出病毒DNA，在感染后312小时后仍能从十二指肠、空肠、盲肠、心脏、肝脏、肾脏，粪便中检出病毒DNA。经滴鼻感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时后能够从心脏、肝脏中检出病毒DNA，感染后24小时能够从胸腺、十二指肠、直肠、空肠、肺脏、心脏、肝脏中检测出病毒DNA，感染后48小时能够从小脑、延髓、胰、盲肠、法氏囊、脾脏、肌肉、骨髓、粪便、血液中检出病毒DNA，在感染后312小时后仍能从胸腺、十二指肠、空肠、盲肠、直肠、心脏、肝脏、肾脏、粪便中检出病毒DNA。三个实验组的鹅在感染后83天均不能从体内检出病毒DNA。经皮下感染后病毒在雏鹅体内的分布速度较经口服和滴鼻感染后病毒在雏鹅体内分布速度快。各种受检组织中心脏和肝脏的检出率最高。经不同途径感染后，病毒均在感染后72小时在机体内各组织的分布最为广泛。本研究的三个实验组(口服，滴鼻，皮下注射)，均未能从大脑、舌头、腺胃、食道中检出病毒DNA。
Primers were designed based on the complete nucleotide sequence of goose parvovirus B strain that zadori subjected to Genebank in 1995,the PCR method for goose parvovirus detedtion was developed.Using this method we can amplify a differential fragment about 441bp from GPV sample ,but we cann't get the same fragment from duck plague virus.Ecoli O1 and O8,MDPV, NGVEV samples.By this method a minimal sample of 0.47pg goose parvovirus nucleotide can be detected.
Using the PCR method developed above,we studied the distribution regularity of virulent goose parvovirs in artifical infected gosling body.The result indicate that there exist difference among the three group(subcutaneously,orally,intranasally).The result of orally infected group: 4 hours after infected ,the GPV nucleotide cann't be detedted from all samples collected; 8 hours afete infected, the GPV nucleotide can be detected from heart and liver;24 hours after infected, the GPV nucleotide can be detedted from thymus,pancreas,duodenum,small intestine,bursa fabricius ,heart,liver,spleen,kidney and blood;48 hours after infected, the GPV nucleotide can be firstly detected from rectum,bone marrow and lung;72 hours after infected , the GPV nucleotide can be detected from cerebellum ,mudulla oblongata,appendix,skeletal muscle,feces.312 hours after infected the GPV nucleotide still can be detected from duodenum,jejunum,rectum,heart,liver,kidney,skeletal muscle,feces.The result of subcutaneously infected group:4 ho
urs after infected, the GPV nucleotide cann't be detedted from all sample collected;8 hours after infected, the GPV nucleotide can be detected from heart and liver;24 hours after infected, the GPV nucleotide can be detected from mudulla oblongata,thymus,pancreas,duodenum,small intestine,appendix,bursa fabricius, heart, liver spleen,lung,kidney,blood,cerebellum,rectum,skeletal muscle,feces;48 hours after infected ,the GPV nucleotide can be firstly detected from bone marrow;312 hours after infected,the GPV nucleotide can still be detected from duodenum, small intestine,appendix,heart,liver,kidney,feces.The result of drop nose:4 hours after infected,the GPV nucleotide cann't be detected from all sample collected,8 hours after infected,the GPV nucleotide can be detected from heart and liver.24 hours after infected,the GPV nucleotide can be detected from thymus,duodenum,rectum,jejunum,lung,cardiac muscle,48 hours after infected GPV nucleotide can be detected from cerebellum, medulla oblongata, Pancreas, cecum, bu
rsa of Fabricius, spleen, Skeletal muscles, bone marrow, feces,blood.312 hours after infected GPV nucleotide can be detected from thymus, duodenum, jejunum, cecum, rectum, cardiac muscle, liver,Kidney,feces.during the period of this studies, we haven't detected the GPV nucleotide from cerebrum,tongue,glandular stomache and gullet.83 days after infection,the virus DNA can't be detected from any tissues of goslings.liver,carciac muscle, jejunum are the most suitable tissues for virus isolation.72 hours or so after infection GPV distribute most widely in gosling.
运用所建立的特异的PCR方法，对人工感染鹅细小病毒后病毒在雏鹅体内的分布规律进行了研究，结果表明：病毒在雏鹅体内的分布规律因病毒感染的途径的不同而存在差异。经口感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时能从心脏、肝脏中检出病毒DNA、感染后24小时能从胸腺、胰腺、十二指肠、小肠、法氏囊、心脏、肝脏、脾脏、肾脏、血液中检出病毒DNA，感染后48小时后能够从直肠、骨髓、肺脏中检出病毒DNA，感染后72小时能从小脑、延髓、盲肠、肌肉、粪便中检出病毒DNA，感染后312小时后仍能从十二指肠、空肠、直肠、心脏、肝脏、肾脏、肌肉、粪便中检出病毒DNA。经皮下注射感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时能够从心脏，肝脏中检测出病毒DNA，感染后24小时能够从延髓、胸腺、胰腺、十二指肠、空肠、盲肠、法氏囊、心脏、肝脏、脾脏、肺脏、肾脏、血液、小脑、直肠、肌肉、粪便中检出病毒DNA，感染后48小时后能从骨髓中检测出病毒DNA，在感染后312小时后仍能从十二指肠、空肠、盲肠、心脏、肝脏、肾脏，粪便中检出病毒DNA。经滴鼻感染组在感染后4小时不能从组织中检出病毒DNA，感染后8小时后能够从心脏、肝脏中检出病毒DNA，感染后24小时能够从胸腺、十二指肠、直肠、空肠、肺脏、心脏、肝脏中检测出病毒DNA，感染后48小时能够从小脑、延髓、胰、盲肠、法氏囊、脾脏、肌肉、骨髓、粪便、血液中检出病毒DNA，在感染后312小时后仍能从胸腺、十二指肠、空肠、盲肠、直肠、心脏、肝脏、肾脏、粪便中检出病毒DNA。三个实验组的鹅在感染后83天均不能从体内检出病毒DNA。经皮下感染后病毒在雏鹅体内的分布速度较经口服和滴鼻感染后病毒在雏鹅体内分布速度快。各种受检组织中心脏和肝脏的检出率最高。经不同途径感染后，病毒均在感染后72小时在机体内各组织的分布最为广泛。本研究的三个实验组(口服，滴鼻，皮下注射)，均未能从大脑、舌头、腺胃、食道中检出病毒DNA。
Primers were designed based on the complete nucleotide sequence of goose parvovirus B strain that zadori subjected to Genebank in 1995,the PCR method for goose parvovirus detedtion was developed.Using this method we can amplify a differential fragment about 441bp from GPV sample ,but we cann't get the same fragment from duck plague virus.Ecoli O1 and O8,MDPV, NGVEV samples.By this method a minimal sample of 0.47pg goose parvovirus nucleotide can be detected.
Using the PCR method developed above,we studied the distribution regularity of virulent goose parvovirs in artifical infected gosling body.The result indicate that there exist difference among the three group(subcutaneously,orally,intranasally).The result of orally infected group: 4 hours after infected ,the GPV nucleotide cann't be detedted from all samples collected; 8 hours afete infected, the GPV nucleotide can be detected from heart and liver;24 hours after infected, the GPV nucleotide can be detedted from thymus,pancreas,duodenum,small intestine,bursa fabricius ,heart,liver,spleen,kidney and blood;48 hours after infected, the GPV nucleotide can be firstly detected from rectum,bone marrow and lung;72 hours after infected , the GPV nucleotide can be detected from cerebellum ,mudulla oblongata,appendix,skeletal muscle,feces.312 hours after infected the GPV nucleotide still can be detected from duodenum,jejunum,rectum,heart,liver,kidney,skeletal muscle,feces.The result of subcutaneously infected group:4 ho
urs after infected, the GPV nucleotide cann't be detedted from all sample collected;8 hours after infected, the GPV nucleotide can be detected from heart and liver;24 hours after infected, the GPV nucleotide can be detected from mudulla oblongata,thymus,pancreas,duodenum,small intestine,appendix,bursa fabricius, heart, liver spleen,lung,kidney,blood,cerebellum,rectum,skeletal muscle,feces;48 hours after infected ,the GPV nucleotide can be firstly detected from bone marrow;312 hours after infected,the GPV nucleotide can still be detected from duodenum, small intestine,appendix,heart,liver,kidney,feces.The result of drop nose:4 hours after infected,the GPV nucleotide cann't be detected from all sample collected,8 hours after infected,the GPV nucleotide can be detected from heart and liver.24 hours after infected,the GPV nucleotide can be detected from thymus,duodenum,rectum,jejunum,lung,cardiac muscle,48 hours after infected GPV nucleotide can be detected from cerebellum, medulla oblongata, Pancreas, cecum, bu
rsa of Fabricius, spleen, Skeletal muscles, bone marrow, feces,blood.312 hours after infected GPV nucleotide can be detected from thymus, duodenum, jejunum, cecum, rectum, cardiac muscle, liver,Kidney,feces.during the period of this studies, we haven't detected the GPV nucleotide from cerebrum,tongue,glandular stomache and gullet.83 days after infection,the virus DNA can't be detected from any tissues of goslings.liver,carciac muscle, jejunum are the most suitable tissues for virus isolation.72 hours or so after infection GPV distribute most widely in gosling.
引文
[1]B．W卡尔尼克主编．禽病学[M](第十版)．高福，苏敬良等译．中国农业出版社，1999，988～995
[2]殷震．刘景华主编．动物病毒学[M](第二版)，北京中国科学出版社，1995：1165～1168．
[3]方定一，小鹅瘟介绍[J]．中国畜牧兽医，1962，8：19～20
[4]Krauss.H.,1965.A disease of goslings. Causing heavy losses[J]. Berlinger Munchener Tieraarztliche Wochenschrift, 78:372～3754
[5]Van Cleef, S.A.M.et al.,1966.A serious disease with an acute course and high ortality in goslings[J]. Tijdschr. Diergeneesk. 91:372～382
[6]Kravchenko, T. M. et al., 1966.Aetiology of a mass disease of goslings[J]. Veterinariya, Moscow, 43(8): 80～90.
[7]Kontrimavichus, L. et al., 1966. Infectious enteritis in goslings [J]. Veterinariya, Moscow, 43(12):34～36.
[8]Derzsy, D. et al., 1966. Aetiology of goose influenza[J]. Magy. Alatorv Lap. 21:385～388.
[9]Wachnik, Z., and J. Novaki 1962. Wirosowe zapaleine watroby u gesiat[J]. Med Weter 18: 344～347
[10]Krauss,H. 1965. Eine Verlustreiche aufzuchtkrankhert bei gansekuker[J].Berl Munch Tieraerztl Wochenschr 78: 372～375.
[11]Angelacev, A. 1966. Exudative septicaemia of goose influenza[J] .Vet Sb Sofia 63: 912
[12]Derzsy, D. 1967. A viral disease of goslings[J]. Acta Vet Hung 17: 443～448
[13]Crighton, G.W., 1970. A disease of goslings[J]. Recueil de medecine Veterinaire, 148(4): 455～472
[14]Schettler, C.H. 1973. Vires hepatitis of geese. Ⅲ. Properties of the causal agent[J]. Avian Pathol 2: 179～193.
[15]Samberg, Y. et al., 1972. A new infectious disease of gosling in Israel[J]. Refuah Veterinarith, 29(1): 29～33
[16]Coudert, M. et al., 1972. Viral disease of goslings[J]. Recueil de medecine Veterinaire, 148 (4): 455～472
[17]Derzsy, D. 1978. A viral disease of goslings In H. Rohrer(ed). Handbuch der Virusinfektionen bei Tieren VI/2[J]. VEB Gustav Fischer Verlag, pp.91～949
[18]方定一，王永坤．小鹅瘟病原体及特异防治的研究[J]．中国农业科学，1981，4：1～8
[19]Gough, R.E., and D.Spackman. 1982. Studies with a duck embryo adapted goose parvovirus [J]. Avian Pathol 11:503～510.
[20]Malkinson, M., B.A.Peleg, R.Nily, and Ekalmar. 1974. The assay of gosling hepatitis virus and antibdy by spermagglutination and spermagglutination-inhibition. Ⅱ. Spermagglutination-inhibition. Avian[J] Pathol 3: 201～209.
[21]郝虹，潘惟均．小鹅瘟病毒核酸链型与结构蛋白分析[J]．病毒学报，1988，4(1)：8～31
[22]郑玉美，李俊宝，周阳生．小鹅瘟病毒核酸型的鉴定[J]．江苏农学院学报，1985，6(4)：45～47
[23]常国权，杨盛华．小鹅瘟病毒的直接透射电镜检测[J]．中国兽医学报．1996，16(1)．93～93
[24]李茂祥，李俊宝，郑玉美．小鹅瘟病毒纯化及其理化特性的研究[J]．病毒学报，1990，6(2)：155～159
[25]Le Gall-Recule, G., and V. Jestin. 1994. Biochemical and genomic characterisatioon of
Muscovy duck parvovirus[J]. Arch Virology 139: 121～131.
[26]佘永建．利用琼脂免疫扩散试验检测小鹅瘟病毒抗原及抗体的研究[J]，畜牧与兽医，1987，5：202～204．
[27]郑玉美．琼脂扩散实验对小鹅瘟病毒检验的研究[J]，畜牧与兽医，1987，4：148～149．
[28]Roszkowski,J.et al. Application of the Imunoperoxidase techniaque for the detection of Derzsy's virus antigen in cell culture and goslings[J]. Avian Pathol. 1982, 11: 571～578.
[29]kardi, V., Szegletes, E.Use of ELISA proedures for the detection of Derzsy's disease virus of goose and of antibodies produced against it[J]. Avian Pathol. 1996, 25: 25～34.
[30]廖德惠，谢镜怀．斑点酶联免疫吸附试验检测小鹅瘟病毒的研究[J]．四川畜牧兽医．1994，(3)．5～6．
[31]邹叔和，李心坦．ABC-ELISA检测小鹅瘟的研究[J]．动物检疫．1992，9(3)．6～7．
[32]潘玉民，董玉平．小鹅瘟免疫荧光诊断方法的研究[J]．中国兽医科技．1990，(10)．6～10．
[33]孙怀昌，廖兴佳．用抗小鹅瘟病毒单抗IgG建立反向间接血凝试验研究[J]．中国畜禽传染病．1989，(2)．44～46．
[34]Malkinson, M. ET AL., 1974. The assay of goslin hepatitis virus and antibody by spermagglutination and SA-inhibition[J], Avain Pathology, 3(3): 201～209.
[35]黄伟，佘永建．应用SPA协同凝集实验快速检测鹅细小病毒．中国畜牧兽医学会禽病研究会第三次代表大会及第五次学术研讨会论文摘要集[M]．1990：辽宁，大连．p128
[36]Gough, R.E., and D.Spackman. 1982. Studies with a duck embryo adapted goose parvovirus[J]. Avian Pathol 11:503～510
[37]段玉友．鹅细小病毒核酸探针的制备和应用[J]．中国畜禽传染病，1993，5：37～39．
[38]谢巧根，徐罗康．小鹅瘟的诊治[J]．养禽与禽病防治．2000，(2)．43～43
[39]王永坤，田慧芳．鹅几种主要传染病的防治[J]．中国禽业导刊．2000，17(17)．11～12
[40]刘玉英 刘娣．如何诊治小鹅瘟[J]．辽宁畜牧兽医．2001，(2)．22～23
[41]刘玉锋，佘锐萍．小鹅瘟的诊断[J]．养禽与禽病防治．2001，(5)．36～36
[42]彭万强 朱治远．小鹅瘟鸭胚化北毒疫苗对雏鹅的免疫研究[J]．中国兽医科技．1992，22(4)．9-11
[43]张英 王玉兰，抗小鹅瘟血清和卵黄抗体制备及推广试验[J]．辽宁畜牧兽医．1991，(4)．14～16
[44]张玉树，熊吉环．抗小鹅瘟血清及卵黄抗体的制备及免疫试验[J]．黑龙江畜牧兽医．1989，(12)．1～3
[45]刘登明，廖德惠．鸭胚化小鹅瘟弱毒疫苗对晚期鹅胚及雏鹅安全性和免疫[J]．中国畜禽传染病．1990，(2)．8～10
[46]黄承锋，朱治远．鹅瘟弱毒株培育研究[J]．畜牧兽医科技．1989，(1)．1～4．
[47]余永建，黄伟．鸭胚化小鹅瘟疫免疫母鹅抗体消长规律的研究[J]．中国兽医杂志．1989，15(10)．49～52．
[48]咸景芳．利用山羊制备抗小鹅瘟血清[J]．现代化农业．1989，(6)．20～21
[49]赖国英 丁新．中药防治小鹅瘟和临床观察[J]．中兽医学杂志．1998，(4)．11～12
[50]薛志成．中西医结合防治小鹅瘟[J]．湖南畜牧兽医．1999，(2)．15～15
[51]黄定余 冯运红．猪白细胞干扰素紧急防治小鹅瘟试验[J]．湖北畜牧兽医．2000，(4)．28～29．
[52]Kisary, J. Interaction in replication between the goose Prvovirus strain B and duck plague herpesvirus [J]. Arch. Virol. 1979,59:81～88.
[53]Deborah. H.Comparative Characterization of Rep Proteins from the Helper-Dependent
Adeno-Associated Virus Type 2 and the Autonomous Goose Parvovims[J]. Journal of Virology, 1999, 73(2); 2930～2937.
[54]Zadori,Z.,R. Stefancsik,T. Rauch,er al. Analysis of the complete nucleotide sequences of goose and muscovy duck parvoviruses indicates common ancestral origin with adenoassociated vires 2 [J].Virology New York. 1995, 212(2): 562～573.
[55]林万明．PCR技术操作和应用指南[M]．北京．人民军医出版社．1995，1～4
[56]富军．小鹅瘟GPV-Scal5弱毒株培育及其特性研究，四川农业大学研究生论文，2002．
[57]Kisary, J.,and D.Derzsy. 1974.A viral disease of goslings. Ⅳ. Characterization of the causal agent in tissue culture systems[J].Acta Vet Hung 24:287～292.
[58]Kontrimavichus, L.M., V.F. Makogon, and V.V. Navrotkii. 1980. Epidemiological,clinical and pathological features of goose viral enteritis[J].Vet Moscow 7: 34～35
[59]Winteroll, G. 1974. Fluorescent antibody studies on goose hepatitis. Proc Goose Dis Symp, Doom Netherlands. pp.65～67.
[60]Bondarenko, A.F. 1982. Improved diagnosis for parvoviral enteritis in geese. (Immunodiffusion test)[J].Arch Exp Vet Med 41:212～221.
[61]汤明，廖德惠．应用改良琼脂免疫扩散试验检测小鹅瘟病毒的研究[J]．中国兽医杂志．1994，20(8)．7～8．
[62]秦爱建，王永坤．免疫酶琼脂扩散试验在小鹅瘟诊断中的应用[J]．中国畜禽传染病，1993，1：30～31．
[63]Have, P., and H.C. Hansen. 1981. Detection of goose parvovirus antibodies by microneutralisation and enzymelinked immunosorbent assay. Proc 7th world Vet poult Assoc, Oslo, Norway, p.60
[64]秦爱建，周阳生，免疫酶斑点法快速检测小鹅瘟病毒[J]．中国兽医科技．1990，(7)．28～29
[65]孙怀昌，李俊宝，朱少旋等．用抗小鹅瘟病毒单抗IgG建立反向间接血凝试验的研究[J]．中国畜禽传染病，1989，2：44～46．
[66]段玉友，崔治中．鹅细小病毒核酸探针的制备及应用[J]．中国畜禽传染病．1993，5：37～39
[67]李新华．应用斑点酶联免疫吸附实验快速诊断小鹅瘟的研究[J]．中国兽医科技．1998．28(1)21～22
[68]余兵，王永坤，朱国强．应用核酸斑点杂交法检测鹅细小病毒(GPV)[J]．中国兽医学报．2002．22(9)453～454．
[69]周复春．PCR技术检测猪伪狂犬病毒及其潜伏感染部位的研究[J]．动物医学进展1997，18(2)22～25．
[70]谢芝勋，Mazhari. Khan, Theodore Girshick.应用PCR检测人工感染鸡体内鸡传染性贫血病毒的动态分布[J]，中国兽医科技1999：29(2)，5～7．
[71]刘培欣，曹殿军，孙建宏．NDV活疫苗及免疫攻毒后组织中病毒分布动态研究[J]，中国预防兽医学报，2000，22，增刊
[72]黄奇昌，程安春，汪铭书．小鹅瘟的研究——弱毒在雏鹅和成年鹅体内分布和排泄[J]．四川农业大学学报1997年15：(1)102～106．
[73]汪铭书，程安春，陈孝跃．鸭病毒性肝炎研究：强毒在雏鸭和成年鸭体内的分布和排泄[J]．中国兽医学报．1997，17(3)：254～257．
[74]任慧英，杨汉春，高云．猪繁殖与呼吸综合征病毒核酸在仔猪呼吸及繁殖系统动态分布的研究[J]，中国兽医杂志，2002．38．(2)．3～4
[75]茂菊，徐福南，洪大蓉．鸡大肝和大脾病抗原在免疫器官的定位和分布[J]，南京农业
大学．2000，23(1)77～80．
[76]张君，由善智．兔出血症病毒在患兔各组织器官内的分布[J]．黑龙江畜牧兽医，1998，12：32-33
[77]林祥梅，赵增连，陈万芳．鸭源流感病毒人工感染鸡的病毒抗原定位动态[J]．中国兽医学报．2001，5，(21)：466～470
[78]许金俊，陶建平，许益民．免疫器官中肾型鸡传染性支气管炎病毒的定位和动态分布[J]，中国预防兽医学报，2001．6(23)：410～413
[79]肖成蕊，梁基．实验感染病毒性关节炎病毒在鸡体内的分布和消长规律的研究[J]．中国兽医杂志，1993，19(10)：8～10
[80]李广兴，任晓峰，杨贵君等．网状内皮组织增殖病病毒感染SPF雏鸡后病毒在各器官内的分布．中国兽医科技．1999，29(11)：6～8．
[81]江国托，王永坤．肾型鸡传染性支气管炎不同脏器中病毒抗原的分布规律[J]．中国兽医杂志．1996，22(6)：8～9
[82]王占森，鹅瘟的研究[J]．中国兽医科技．1993．23(3)：7～9
[2]殷震．刘景华主编．动物病毒学[M](第二版)，北京中国科学出版社，1995：1165～1168．
[3]方定一，小鹅瘟介绍[J]．中国畜牧兽医，1962，8：19～20
[4]Krauss.H.,1965.A disease of goslings. Causing heavy losses[J]. Berlinger Munchener Tieraarztliche Wochenschrift, 78:372～3754
[5]Van Cleef, S.A.M.et al.,1966.A serious disease with an acute course and high ortality in goslings[J]. Tijdschr. Diergeneesk. 91:372～382
[6]Kravchenko, T. M. et al., 1966.Aetiology of a mass disease of goslings[J]. Veterinariya, Moscow, 43(8): 80～90.
[7]Kontrimavichus, L. et al., 1966. Infectious enteritis in goslings [J]. Veterinariya, Moscow, 43(12):34～36.
[8]Derzsy, D. et al., 1966. Aetiology of goose influenza[J]. Magy. Alatorv Lap. 21:385～388.
[9]Wachnik, Z., and J. Novaki 1962. Wirosowe zapaleine watroby u gesiat[J]. Med Weter 18: 344～347
[10]Krauss,H. 1965. Eine Verlustreiche aufzuchtkrankhert bei gansekuker[J].Berl Munch Tieraerztl Wochenschr 78: 372～375.
[11]Angelacev, A. 1966. Exudative septicaemia of goose influenza[J] .Vet Sb Sofia 63: 912
[12]Derzsy, D. 1967. A viral disease of goslings[J]. Acta Vet Hung 17: 443～448
[13]Crighton, G.W., 1970. A disease of goslings[J]. Recueil de medecine Veterinaire, 148(4): 455～472
[14]Schettler, C.H. 1973. Vires hepatitis of geese. Ⅲ. Properties of the causal agent[J]. Avian Pathol 2: 179～193.
[15]Samberg, Y. et al., 1972. A new infectious disease of gosling in Israel[J]. Refuah Veterinarith, 29(1): 29～33
[16]Coudert, M. et al., 1972. Viral disease of goslings[J]. Recueil de medecine Veterinaire, 148 (4): 455～472
[17]Derzsy, D. 1978. A viral disease of goslings In H. Rohrer(ed). Handbuch der Virusinfektionen bei Tieren VI/2[J]. VEB Gustav Fischer Verlag, pp.91～949
[18]方定一，王永坤．小鹅瘟病原体及特异防治的研究[J]．中国农业科学，1981，4：1～8
[19]Gough, R.E., and D.Spackman. 1982. Studies with a duck embryo adapted goose parvovirus [J]. Avian Pathol 11:503～510.
[20]Malkinson, M., B.A.Peleg, R.Nily, and Ekalmar. 1974. The assay of gosling hepatitis virus and antibdy by spermagglutination and spermagglutination-inhibition. Ⅱ. Spermagglutination-inhibition. Avian[J] Pathol 3: 201～209.
[21]郝虹，潘惟均．小鹅瘟病毒核酸链型与结构蛋白分析[J]．病毒学报，1988，4(1)：8～31
[22]郑玉美，李俊宝，周阳生．小鹅瘟病毒核酸型的鉴定[J]．江苏农学院学报，1985，6(4)：45～47
[23]常国权，杨盛华．小鹅瘟病毒的直接透射电镜检测[J]．中国兽医学报．1996，16(1)．93～93
[24]李茂祥，李俊宝，郑玉美．小鹅瘟病毒纯化及其理化特性的研究[J]．病毒学报，1990，6(2)：155～159
[25]Le Gall-Recule, G., and V. Jestin. 1994. Biochemical and genomic characterisatioon of
Muscovy duck parvovirus[J]. Arch Virology 139: 121～131.
[26]佘永建．利用琼脂免疫扩散试验检测小鹅瘟病毒抗原及抗体的研究[J]，畜牧与兽医，1987，5：202～204．
[27]郑玉美．琼脂扩散实验对小鹅瘟病毒检验的研究[J]，畜牧与兽医，1987，4：148～149．
[28]Roszkowski,J.et al. Application of the Imunoperoxidase techniaque for the detection of Derzsy's virus antigen in cell culture and goslings[J]. Avian Pathol. 1982, 11: 571～578.
[29]kardi, V., Szegletes, E.Use of ELISA proedures for the detection of Derzsy's disease virus of goose and of antibodies produced against it[J]. Avian Pathol. 1996, 25: 25～34.
[30]廖德惠，谢镜怀．斑点酶联免疫吸附试验检测小鹅瘟病毒的研究[J]．四川畜牧兽医．1994，(3)．5～6．
[31]邹叔和，李心坦．ABC-ELISA检测小鹅瘟的研究[J]．动物检疫．1992，9(3)．6～7．
[32]潘玉民，董玉平．小鹅瘟免疫荧光诊断方法的研究[J]．中国兽医科技．1990，(10)．6～10．
[33]孙怀昌，廖兴佳．用抗小鹅瘟病毒单抗IgG建立反向间接血凝试验研究[J]．中国畜禽传染病．1989，(2)．44～46．
[34]Malkinson, M. ET AL., 1974. The assay of goslin hepatitis virus and antibody by spermagglutination and SA-inhibition[J], Avain Pathology, 3(3): 201～209.
[35]黄伟，佘永建．应用SPA协同凝集实验快速检测鹅细小病毒．中国畜牧兽医学会禽病研究会第三次代表大会及第五次学术研讨会论文摘要集[M]．1990：辽宁，大连．p128
[36]Gough, R.E., and D.Spackman. 1982. Studies with a duck embryo adapted goose parvovirus[J]. Avian Pathol 11:503～510
[37]段玉友．鹅细小病毒核酸探针的制备和应用[J]．中国畜禽传染病，1993，5：37～39．
[38]谢巧根，徐罗康．小鹅瘟的诊治[J]．养禽与禽病防治．2000，(2)．43～43
[39]王永坤，田慧芳．鹅几种主要传染病的防治[J]．中国禽业导刊．2000，17(17)．11～12
[40]刘玉英 刘娣．如何诊治小鹅瘟[J]．辽宁畜牧兽医．2001，(2)．22～23
[41]刘玉锋，佘锐萍．小鹅瘟的诊断[J]．养禽与禽病防治．2001，(5)．36～36
[42]彭万强 朱治远．小鹅瘟鸭胚化北毒疫苗对雏鹅的免疫研究[J]．中国兽医科技．1992，22(4)．9-11
[43]张英 王玉兰，抗小鹅瘟血清和卵黄抗体制备及推广试验[J]．辽宁畜牧兽医．1991，(4)．14～16
[44]张玉树，熊吉环．抗小鹅瘟血清及卵黄抗体的制备及免疫试验[J]．黑龙江畜牧兽医．1989，(12)．1～3
[45]刘登明，廖德惠．鸭胚化小鹅瘟弱毒疫苗对晚期鹅胚及雏鹅安全性和免疫[J]．中国畜禽传染病．1990，(2)．8～10
[46]黄承锋，朱治远．鹅瘟弱毒株培育研究[J]．畜牧兽医科技．1989，(1)．1～4．
[47]余永建，黄伟．鸭胚化小鹅瘟疫免疫母鹅抗体消长规律的研究[J]．中国兽医杂志．1989，15(10)．49～52．
[48]咸景芳．利用山羊制备抗小鹅瘟血清[J]．现代化农业．1989，(6)．20～21
[49]赖国英 丁新．中药防治小鹅瘟和临床观察[J]．中兽医学杂志．1998，(4)．11～12
[50]薛志成．中西医结合防治小鹅瘟[J]．湖南畜牧兽医．1999，(2)．15～15
[51]黄定余 冯运红．猪白细胞干扰素紧急防治小鹅瘟试验[J]．湖北畜牧兽医．2000，(4)．28～29．
[52]Kisary, J. Interaction in replication between the goose Prvovirus strain B and duck plague herpesvirus [J]. Arch. Virol. 1979,59:81～88.
[53]Deborah. H.Comparative Characterization of Rep Proteins from the Helper-Dependent
Adeno-Associated Virus Type 2 and the Autonomous Goose Parvovims[J]. Journal of Virology, 1999, 73(2); 2930～2937.
[54]Zadori,Z.,R. Stefancsik,T. Rauch,er al. Analysis of the complete nucleotide sequences of goose and muscovy duck parvoviruses indicates common ancestral origin with adenoassociated vires 2 [J].Virology New York. 1995, 212(2): 562～573.
[55]林万明．PCR技术操作和应用指南[M]．北京．人民军医出版社．1995，1～4
[56]富军．小鹅瘟GPV-Scal5弱毒株培育及其特性研究，四川农业大学研究生论文，2002．
[57]Kisary, J.,and D.Derzsy. 1974.A viral disease of goslings. Ⅳ. Characterization of the causal agent in tissue culture systems[J].Acta Vet Hung 24:287～292.
[58]Kontrimavichus, L.M., V.F. Makogon, and V.V. Navrotkii. 1980. Epidemiological,clinical and pathological features of goose viral enteritis[J].Vet Moscow 7: 34～35
[59]Winteroll, G. 1974. Fluorescent antibody studies on goose hepatitis. Proc Goose Dis Symp, Doom Netherlands. pp.65～67.
[60]Bondarenko, A.F. 1982. Improved diagnosis for parvoviral enteritis in geese. (Immunodiffusion test)[J].Arch Exp Vet Med 41:212～221.
[61]汤明，廖德惠．应用改良琼脂免疫扩散试验检测小鹅瘟病毒的研究[J]．中国兽医杂志．1994，20(8)．7～8．
[62]秦爱建，王永坤．免疫酶琼脂扩散试验在小鹅瘟诊断中的应用[J]．中国畜禽传染病，1993，1：30～31．
[63]Have, P., and H.C. Hansen. 1981. Detection of goose parvovirus antibodies by microneutralisation and enzymelinked immunosorbent assay. Proc 7th world Vet poult Assoc, Oslo, Norway, p.60
[64]秦爱建，周阳生，免疫酶斑点法快速检测小鹅瘟病毒[J]．中国兽医科技．1990，(7)．28～29
[65]孙怀昌，李俊宝，朱少旋等．用抗小鹅瘟病毒单抗IgG建立反向间接血凝试验的研究[J]．中国畜禽传染病，1989，2：44～46．
[66]段玉友，崔治中．鹅细小病毒核酸探针的制备及应用[J]．中国畜禽传染病．1993，5：37～39
[67]李新华．应用斑点酶联免疫吸附实验快速诊断小鹅瘟的研究[J]．中国兽医科技．1998．28(1)21～22
[68]余兵，王永坤，朱国强．应用核酸斑点杂交法检测鹅细小病毒(GPV)[J]．中国兽医学报．2002．22(9)453～454．
[69]周复春．PCR技术检测猪伪狂犬病毒及其潜伏感染部位的研究[J]．动物医学进展1997，18(2)22～25．
[70]谢芝勋，Mazhari. Khan, Theodore Girshick.应用PCR检测人工感染鸡体内鸡传染性贫血病毒的动态分布[J]，中国兽医科技1999：29(2)，5～7．
[71]刘培欣，曹殿军，孙建宏．NDV活疫苗及免疫攻毒后组织中病毒分布动态研究[J]，中国预防兽医学报，2000，22，增刊
[72]黄奇昌，程安春，汪铭书．小鹅瘟的研究——弱毒在雏鹅和成年鹅体内分布和排泄[J]．四川农业大学学报1997年15：(1)102～106．
[73]汪铭书，程安春，陈孝跃．鸭病毒性肝炎研究：强毒在雏鸭和成年鸭体内的分布和排泄[J]．中国兽医学报．1997，17(3)：254～257．
[74]任慧英，杨汉春，高云．猪繁殖与呼吸综合征病毒核酸在仔猪呼吸及繁殖系统动态分布的研究[J]，中国兽医杂志，2002．38．(2)．3～4
[75]茂菊，徐福南，洪大蓉．鸡大肝和大脾病抗原在免疫器官的定位和分布[J]，南京农业
大学．2000，23(1)77～80．
[76]张君，由善智．兔出血症病毒在患兔各组织器官内的分布[J]．黑龙江畜牧兽医，1998，12：32-33
[77]林祥梅，赵增连，陈万芳．鸭源流感病毒人工感染鸡的病毒抗原定位动态[J]．中国兽医学报．2001，5，(21)：466～470
[78]许金俊，陶建平，许益民．免疫器官中肾型鸡传染性支气管炎病毒的定位和动态分布[J]，中国预防兽医学报，2001．6(23)：410～413
[79]肖成蕊，梁基．实验感染病毒性关节炎病毒在鸡体内的分布和消长规律的研究[J]．中国兽医杂志，1993，19(10)：8～10
[80]李广兴，任晓峰，杨贵君等．网状内皮组织增殖病病毒感染SPF雏鸡后病毒在各器官内的分布．中国兽医科技．1999，29(11)：6～8．
[81]江国托，王永坤．肾型鸡传染性支气管炎不同脏器中病毒抗原的分布规律[J]．中国兽医杂志．1996，22(6)：8～9
[82]王占森，鹅瘟的研究[J]．中国兽医科技．1993．23(3)：7～9