犬细小病毒病临床诊断方法的筛选及新治疗手段的研究
摘要
对上海市犬细小病毒病进行了发病率调查,同时进行了治疗手段的调查。设计了聚合酶链式反应,免疫胶体金试纸条,酶联免疫吸附试验,血凝试验四种方法诊断犬细小病毒病。并采用单克隆抗体、干扰素、单克隆抗体干扰素相结合等综合治疗措施对该病进行了分组治疗。目的是筛选出当前最佳实验室诊断方法,并选择最适合的治疗措施。结果表明,春季犬细小病毒病在上海市的发病率高,占门诊比例的19.2%以上,应用PCR诊断诊断犬细小病毒病的准确率达95.7%,而且成本低,敏感性,特异性强,完全可以应用于实验室和临床诊断中。实验应用的综合治疗措施对犬细小病毒病早期的治愈率达84%,中期的治愈率达45%,晚期的治愈率为9%以上,是一种行之有效的治疗措施。
The morbidity of Canine parvovirus disease in shanghai was investigated , and the diagonostic method of Polymerase Chain Reaction test PCR for detecting canine parvovirus was used and the treatment method of CPV mcAb was studied .The results showed that the morbidity of canine parvovirus disease was more than 19.2% ,and highest in spring in shanghai aera. The accuracy of the diagnosis using PCR was 95.8%, at the same time, the way was cheap,sensible,specific. In early, metaphase and advanced stage.The cure rate for the canine parvovirus disease by using the treatment method of combination of Chines traditional and western medicine were 84%,45%,9%,respectively. It can be concluded that the diagonostic and treatment are effective,and can be used in practice .
The morbidity of Canine parvovirus disease in shanghai was investigated , and the diagonostic method of Polymerase Chain Reaction test PCR for detecting canine parvovirus was used and the treatment method of CPV mcAb was studied .The results showed that the morbidity of canine parvovirus disease was more than 19.2% ,and highest in spring in shanghai aera. The accuracy of the diagnosis using PCR was 95.8%, at the same time, the way was cheap,sensible,specific. In early, metaphase and advanced stage.The cure rate for the canine parvovirus disease by using the treatment method of combination of Chines traditional and western medicine were 84%,45%,9%,respectively. It can be concluded that the diagonostic and treatment are effective,and can be used in practice .
引文
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[4]王瑞生,吴玉坤.犬出血性肠炎研究近况.中国畜禽传染病,1993,(3):61—62
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[21].Mochizuki M, San Gabriel MC, Nakatani H. Comparison of polymerase chain reaction with virus isolation and haemagglutination assays for the detection of canine parvoviruses in faecal specimens. [J] Vet Sci 1993,55(1):60-63.
[22] Hirasawa T, Kaneshige T, Mikazuki K. Sensitive detection of canine parvovirus DNA by the nested polymerase chain reaction.[J] Vet Microbiol , 1994 ,41(1-2):135-45
[23].Schunc B, Kraft W Truyen U. A simple touch-down polymerase chain reaction for the detection of canine parvovirus and feline panleukopenia virus in feces. Virol Methods ,1995,55(3):427-433
[24] Schwab KJ, Neill FH, Le Guyader F. Development of a reverse transcription-PCR-DNA enzyme immunoassay for detection of "Norwalk-like" virusesand hepatitis A virus in stool and shellfish .Appl Environ Microbiol 2001 ,67(2):742-9.
[25] Kato N, OU CY, Kato H. Detection of toxigenic Clostridium difficile in stool specimens by thepolymerase chain reaction.[J] Infect Dis,1993,167(2):455-458
[26] Widjojoatmodjo MN, Fluit AC, Torensma R. The magnetic immune polymerase chain reaction assay for direct detection of salmonellae in fecal samples. Clin Microbiol ,1992 ,30(12):3195-31999
[27] Saiki RK, Scharf S, FaloonaFEnzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia [J]Science,1985 ,230 (4732): 1350-1354
[28] Mullis K,Faloona FScharf S. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction.[J] Cold Spring Harb Symp Quant Biol, 1986,51 (1):263-73.
[29] Saiki RK, Gelfand DH, Stoffel S.Primer-directed enzymatic amplification of DNA with athermostable DNA polymerase.Science. 1988 ,29:487-491.
[30] 丁振若,苏明权.临床PCR基因诊断技术.世界图书出版西安公司,1998, 1—2.
[31] Belak S ,Thoren P .Molecular diagnosis of animal diseases :some ex- peiences over the past decade. Expert Rev Mol Diagn,2001,(1):434-443.
[32] Jungkind D .Automaiton of laboratory testing for in factious diseases using the polymerase chain reaction—our past, our present ,our future.Clin Virol,2001,20(1-2):l-6.
[33] Claustres M, Kjellberg P, Desgeorges M. Detection of deletions by the amplification of exons (multiplex PCR) in Duchenne muscular dystrophy. [J] Genet Hum,1989,37(3):251-257.
[34] Erlich HA,Gelfand D,Sninsky JJ. Recent advances in the polymerase chain reaction.Science. 1991, 252:1643-1651
[35]SanoTSmith CL, Cantor CR. Immune-PCR: very sensitive antigen detection by means of specific antibody-DNA conjugates.Science,1992,2:120-122
[36] Haase AT, Retzel EF, Staskus KA. Amplification and detection of lentiviral DNA inside cells.[J] ProcNatl Acad Sci U S A ,1990 ,87(13):4971-4975
[37] Murphy LD, Herzog CE, RudickJB.Use of the polymerase chain reaction in the quantitation ofmdr-1 gene expression. Biochemistry ,1990 ,29(45):10351-10356.
[38] 刘宏伟.中国畜禽传染病,1994(6):10—13
[39]Chandler J,Robinson N,Whiting K.Handling false signals in gold— based rapid tests[J].IVD Technology ,2001,7(2):34—45 [4O]Cavalli A,Bozzo G,Decaro N,et a1.Characterization of a canine parvovirus strain isolated from all adult dog[J].New Microbiology, 2001,24:239—242
[41]Cesario C,Decaro N,Campolo M,et a1.Canine parvovirus infection:Which diagnostic test for virus[J].Journal of Virological Methods,2005,126:179—185
[2]. 殷震,刘景华主编.动物病毒学.北京:科学出版社,2000, 1113-1116
[3]. Appel MJ , Scot FW, Carmichael LE. Isolation and immunisation studies of a canine parco-like virus from dogs with haemorrhagic enteritis.[J] Vet Ree ,1979 ,25 (8): 156
[4]王瑞生,吴玉坤.犬出血性肠炎研究近况.中国畜禽传染病,1993,(3):61—62
[5] 王培君,邓树军,慈勤珍,等.中西医结合治疗犬细小病毒病.中兽医医药杂志,2001,(4):31-32.
[6] 俞华英,蒋岩.犬细小病毒的诊治.畜牧兽医杂志2OO2,21(5):39
[7] 叶新华,李兴,王伟.犬细小病毒病的鉴别诊断[J] . 黑龙江畜牧兽医,2002,(5):62.
[8]Annamaria,Pratolli,Alessandra,Cavalli,Vito,Martellaeta1Caninearvovirus(CPV)vaccination:comparison of neutralizing antibody responses in pups after inoculation with CPV2 Or CPV2b modified live virus vaccine.vaccine[J].Clinical and Doagnostic laboratory immunology,2001,5:6l2-6l5.
[9] . Mathys A, Mueller R, Pedersen NC.comparison of hemaggulutiantion and competitiveenzymelinked immunosibent assay procedures for detecting canine parvovirus in feces. Vet Res ,1983,44(1):152-154
[10].Flower RI,Wilcox GE, Robinson WT. Antigenic diferences between canine parvovirus and feline panleucopenia virus. Vet Rec,1980,107(11):254-256.
[11] 吴金石,周建群.运用ELISA迅速检测犬细小病毒抗体方法的建立与均衡性研究[J].微生物杂志,2OO2,22(4):14—16.
[12] Rimmelzwaan GF,Juntti N,klingeborn B,et al.Evaluation of enzyme-linked immunosorbent assays based on monoclonal antibodies forthe serology and antigen detection in cltnine parvovirus infections[J].Vet Q,1990,12(1):14-20.
[13] 马国红.犬细小病毒病的诊治[J].河北畜牧兽医,2OO2,l8 (4):38.
[14] 李六金,姜焕宏,李成,等.免疫金电镜技术检测犬细小病毒免疫球蛋白[J].中国动物检疫,2OOO,17(8):27-28.
[15] Banja-BK,Sahoo-N,Panda-HK,et al.Epizootiological status of canine viral hae mnorhagic gastro-enteritis in Bhubaneswar City [J].Indian Veterinary Journal,2OO2,79(8):850-851.
[16] Rimmelzwaan GF,Groen J,Egberink H,et a1. The use of enzymelinked immunosorbent assay systems for serology and antigendetection in parvovirus,coronavirus and rotavirus infections in dogs in The Netherland [J].Vet Microbiol,1991,26(1-2):25-40,
[17] Subhashini—CR,Meerarani-S,Ramadass- P,et a1.Polymerase reaction and latex agglutination test for detection of canine parvovirus infection[J].Indian Journal of Virology,1997,13(1):65-68.
[18] Rypul K,Chmielewski R,Smielewska-Los E,et a1.Phylogenetic similarity of the canine parvoviruswild type isolates on the basis of VP1 /VP2 gene fragment sequence analysis.[J] Vet Med B Infect Dis VetPublic Health,2OO2,49(3):142—145.
[19] Martinello F,Galuppo F,Ostanello F,et a1.Detection of canine parvovirus in wolves from Italy[J].J Wildl Dis,1997,33(3):628- 631.
[20] Harasawa R, Yoshida M, Kataoka Y. Detection by PCR of genomic markers in canine parvovirus and feline panleukopenia virus.[J] C R Seances Soc Biol Fil 1993,187(4):554-60
[21].Mochizuki M, San Gabriel MC, Nakatani H. Comparison of polymerase chain reaction with virus isolation and haemagglutination assays for the detection of canine parvoviruses in faecal specimens. [J] Vet Sci 1993,55(1):60-63.
[22] Hirasawa T, Kaneshige T, Mikazuki K. Sensitive detection of canine parvovirus DNA by the nested polymerase chain reaction.[J] Vet Microbiol , 1994 ,41(1-2):135-45
[23].Schunc B, Kraft W Truyen U. A simple touch-down polymerase chain reaction for the detection of canine parvovirus and feline panleukopenia virus in feces. Virol Methods ,1995,55(3):427-433
[24] Schwab KJ, Neill FH, Le Guyader F. Development of a reverse transcription-PCR-DNA enzyme immunoassay for detection of "Norwalk-like" virusesand hepatitis A virus in stool and shellfish .Appl Environ Microbiol 2001 ,67(2):742-9.
[25] Kato N, OU CY, Kato H. Detection of toxigenic Clostridium difficile in stool specimens by thepolymerase chain reaction.[J] Infect Dis,1993,167(2):455-458
[26] Widjojoatmodjo MN, Fluit AC, Torensma R. The magnetic immune polymerase chain reaction assay for direct detection of salmonellae in fecal samples. Clin Microbiol ,1992 ,30(12):3195-31999
[27] Saiki RK, Scharf S, FaloonaFEnzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia [J]Science,1985 ,230 (4732): 1350-1354
[28] Mullis K,Faloona FScharf S. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction.[J] Cold Spring Harb Symp Quant Biol, 1986,51 (1):263-73.
[29] Saiki RK, Gelfand DH, Stoffel S.Primer-directed enzymatic amplification of DNA with athermostable DNA polymerase.Science. 1988 ,29:487-491.
[30] 丁振若,苏明权.临床PCR基因诊断技术.世界图书出版西安公司,1998, 1—2.
[31] Belak S ,Thoren P .Molecular diagnosis of animal diseases :some ex- peiences over the past decade. Expert Rev Mol Diagn,2001,(1):434-443.
[32] Jungkind D .Automaiton of laboratory testing for in factious diseases using the polymerase chain reaction—our past, our present ,our future.Clin Virol,2001,20(1-2):l-6.
[33] Claustres M, Kjellberg P, Desgeorges M. Detection of deletions by the amplification of exons (multiplex PCR) in Duchenne muscular dystrophy. [J] Genet Hum,1989,37(3):251-257.
[34] Erlich HA,Gelfand D,Sninsky JJ. Recent advances in the polymerase chain reaction.Science. 1991, 252:1643-1651
[35]SanoTSmith CL, Cantor CR. Immune-PCR: very sensitive antigen detection by means of specific antibody-DNA conjugates.Science,1992,2:120-122
[36] Haase AT, Retzel EF, Staskus KA. Amplification and detection of lentiviral DNA inside cells.[J] ProcNatl Acad Sci U S A ,1990 ,87(13):4971-4975
[37] Murphy LD, Herzog CE, RudickJB.Use of the polymerase chain reaction in the quantitation ofmdr-1 gene expression. Biochemistry ,1990 ,29(45):10351-10356.
[38] 刘宏伟.中国畜禽传染病,1994(6):10—13
[39]Chandler J,Robinson N,Whiting K.Handling false signals in gold— based rapid tests[J].IVD Technology ,2001,7(2):34—45 [4O]Cavalli A,Bozzo G,Decaro N,et a1.Characterization of a canine parvovirus strain isolated from all adult dog[J].New Microbiology, 2001,24:239—242
[41]Cesario C,Decaro N,Campolo M,et a1.Canine parvovirus infection:Which diagnostic test for virus[J].Journal of Virological Methods,2005,126:179—185