水貂血液阿留申病病毒及其抗体的检测与分析
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摘要
为了确定鉴定阿留申病的适宜方法,尤其在阿留申病毒高感染貂场,本试验联合应用对流免疫电泳(CIEP)和PCR方法,对1个貂场连续2年于11~12月间检测1次貂血液的阿留申病毒及其抗体。用CIEP检测产仔(断乳成活数≥5只/窝)母貂及其仔貂。通过比较貂群的检测结果来分析2种检测方法鉴别阿留申病貂的适用性。2次检测结果显示各貂群CIEP阳性率为60%~97%,PCR阳性率为20%~63%,水貂的CIEP与PCR检测阳性结果不完全匹配。根据CIEP和PCR联合检测结果,貂群被分为4个群体CIEP+PCR+、CIEP-PCR-、CIEP-PCR+和CIEP+PCR-。2次检测的CIEP+PCR-貂在当年母貂中分别占48%、39%,在老母貂中分别占43%、77%,在公貂中分别占21%、51%。2次检测的CIEP+PCR-貂在貂群所占比例大,而CIEP-PCR-和CIEP-PCR+貂在貂群中所占比例小。公貂、老母貂经PCR阳性淘汰间隔1年再次检测,CIEP+PCR-貂在群体中比例明显提高。产仔2~4年龄母貂有72%~80%CIEP+貂,而其60日龄仔貂仅有11%~16%CIEP+貂。结果分析指出,中国阿留申病病毒高感染率养殖貂群不适宜单纯采用CIEP检测淘汰阳性貂净化阿留申病。发现每年11至12月用CIEP和PCR联合检测貂血样,易检出CIEP+PCR-的潜在阿留申病毒感染康复貂。
To determine the suitability of the current methods for the diagnosis of aleutian disease in a mink ranch with high seroprevalence of aleutian disease virus.A combination of counter-immune electrophoresis(CIEP)and PCR tests were used to detected aleutian disease virus(ADV)and the antibody to ADV from mink circulating blood between November and December once a year for two consecutive years in a mink ranch.The dams were selected together with their kits based on the litter size,(≥5healthy kits per litter),and CIEP was used to detect the antibody to ADV.The results obtained from different mink herds were analyzed to determine whether PCR and CIEP tests is suitable for the diagnosis of Aleutian disease in the special period.Testing results showed that the positive percentages in a mink herd ranged from 60%to 88%in CIEP test and from 31%to 63%in PCR test.CIEP positive mink did not completely match those in positive PCR.The experimental minks could be divided into four groups based on the CIEP and PCR results,which were as follows:CIEP+PCR+,CIEP-PCR-,CIEP-PCR+ and CIEP+PCR-.In the two detections,percentage for CIEP + PCR-was 48% and 39%in the yearling mink,21% and 51%in sires,43% and 77%in old dams.The results showed there was a higher proportion for CIEP+PCR-,but lower proportion for CIEP-PCR-or CIEP-PCR+.Percentage for CIEP + PCRwas significantly increased 34%(77%,43%)in old dams,and(30%,51%)in sires(through culling PCR positive mink.Percentage for positive CIEP varied from 72%to 80%in the two-year-old or older dams with good calving performance,while it varied from 11%to 16%in their sixty-day-old kits.The results indicated that long-term breeding mink herd had high seroprevalence of Aleutian disease virus,it is inappropriate for the elimination of Aleutian disease to cull the positive mink only by CIEP.We found that between November and December,the combination of CIEP and PCR tests is appropriate to screen minks for CIEP+ PCR-using circulating blood,which may be recovery from ADV hiddent infection.
To determine the suitability of the current methods for the diagnosis of aleutian disease in a mink ranch with high seroprevalence of aleutian disease virus.A combination of counter-immune electrophoresis(CIEP)and PCR tests were used to detected aleutian disease virus(ADV)and the antibody to ADV from mink circulating blood between November and December once a year for two consecutive years in a mink ranch.The dams were selected together with their kits based on the litter size,(≥5healthy kits per litter),and CIEP was used to detect the antibody to ADV.The results obtained from different mink herds were analyzed to determine whether PCR and CIEP tests is suitable for the diagnosis of Aleutian disease in the special period.Testing results showed that the positive percentages in a mink herd ranged from 60%to 88%in CIEP test and from 31%to 63%in PCR test.CIEP positive mink did not completely match those in positive PCR.The experimental minks could be divided into four groups based on the CIEP and PCR results,which were as follows:CIEP+PCR+,CIEP-PCR-,CIEP-PCR+ and CIEP+PCR-.In the two detections,percentage for CIEP + PCR-was 48% and 39%in the yearling mink,21% and 51%in sires,43% and 77%in old dams.The results showed there was a higher proportion for CIEP+PCR-,but lower proportion for CIEP-PCR-or CIEP-PCR+.Percentage for CIEP + PCRwas significantly increased 34%(77%,43%)in old dams,and(30%,51%)in sires(through culling PCR positive mink.Percentage for positive CIEP varied from 72%to 80%in the two-year-old or older dams with good calving performance,while it varied from 11%to 16%in their sixty-day-old kits.The results indicated that long-term breeding mink herd had high seroprevalence of Aleutian disease virus,it is inappropriate for the elimination of Aleutian disease to cull the positive mink only by CIEP.We found that between November and December,the combination of CIEP and PCR tests is appropriate to screen minks for CIEP+ PCR-using circulating blood,which may be recovery from ADV hiddent infection.
引文
[1]Porter D D,Larsen A E,Porter H G.The pathogenesis of aleutian disease of mink i.in vivo viral replication and the host antibody response to viral antigen[J].J Exp Med,1969,130(3):575-593.
[2]Alex S,BloomM E,WolfinbargerJ.Evidence of restricted viral replication in adult mink infected with Aleutian disease of mink parvovirus[J]J Virol,1988,62(5):1495-1507.
[3]Hadlow W J,Race R E,Kennedy R C.Comparative pathogenicity of four strains of Aleutian disease virus for pastel and sapphire mink[J].Infect Immun,1983,41(3):1016-1023.
[4]Hansen M,Lund E.Pregnancy rate and foetal mortality in Aleutian disease virus infected mink[J].Acta Vet Scand,1988,29(2):271-272.
[5]Padgett G A,GorhamJ R,Henson J B.Epizootiologic studies of Aleutian disease.I.Transplacental transmission of the virus[J].J Infect Dis,1967,117(1):35-38.
[6]王兰坡.应用对流免疫电泳检疫水貂阿留申病的报告[J].经济动物学报,1991,2(7):19-21.
[7]Sang Y,M J,Hou Z,et al.Phylogenetic analysis of the VP2gene of Aleutian mink disease parvoviruses isolated from 2009to 2011in China[J].Virus Genes,2012,45(1):31-37.
[8]华育平,马建.阿留申病细小病毒的分离及VP2基因遗传衍生分析[J].畜牧兽医学报,2005,36(9):960-963.
[9]Henson J,GorhamJ,Leader R.A field test for Aleutian disease[J].National Fur News,1962,34:8-9.
[10]Cho H J,Greenfield J.Eradication of Aleutian disease of mink by eliminating positive counterimmunoelectrophoresis test reactors[J].J Clin Microbiol,1978,7(1):18-22.
[11]Bloom M E,OieI K L,Wolfinbarger J,et al.Evaluation of the polymerase chain reaction(pcr)as a tool for diagnosing infections with the Aleutian mink disease parvovirus[J].Scientifur,1997,21(2):141-146.
[12]Cepica A,IwamotoT.Field evaluation of CIEP and PCR detection/removal control methods of Aleutian mink disease(AD)in Canada[C]//Larsen P F,Mller S H,Clausen T,et al.Proceedings of the Xth International scientific congress in fur animal production.Wageningen Academic Publishers,2012:196-205.
[13]Jackson M K,Ellis L C,Morrey J D,et al.Progression of Aleutian disease in natural and experimentally induced infections of mink[J].Am J Vet Res,1996,57(12):1753-1758.
[14]Hadlow W J,RaceR E,Kenneedy R C.Royal pastel mink respond variously to inoculation with Aleutian disease virus of low virulence[J].J Virol,1984,50(1):38-41.
[15]Larsen A E,Porte D D.Pathogenesis of aleutian disease of mink:identification of nonpersistent infections[J].Infect Immun,1975,11(1):92-94.
[16]As S H,Depauli F J,Wright P,et al.Characteristics of inapparent Aleutian disease virus infection in mink[J].Res Vet Sci,1978,24(2):200-204.
[17]An S H,IngaramD G.Detection of inapparent Aleutian disease virus infection in mink[J].Am J Vet Res,1977,38(10):1619-1624.
[18]籍玉林,曲维江,赵元楷,等.阿留申病抗体消长规律的研究[J].畜牧兽医学报,1995,26(1):42-46.
[19]黄培堂,王嘉玺,朱厚咄译.分子克隆实验指南[M].3版,北京:科学出版社,2002,467-469.
[20]赖平安,张鹤晓,刘艳华.水貂阿留申病对流免疫电泳操作规程GB/T 6682[S]//中华人民共和国北京出入境检验检疫局,中国;中华人民共和国国家质量监督检验检疫总局,2003-08-18.
[21]J.P K.Calculation for the chi-square test:An interactive calculation tool for chi-square tests of goodness of fit and independence[E].2001,April.
[22]Li Y,HuangJ,Jia Y,et al.Genetic characterization of Aleutian mink disease viruses isolated in China[J].Virus genes,2012,45(1):24-30.
[23]Cepica A,Zargar A,Mohamed B,et al.In vivo MALDI-TOF markers for early detection of Aleutian disease(AD)among the AD virus infected mink[C]//LarsenP F,Ller S H,Clausen T,et al.Proceedings of the Xth International Scientific Congress in fur animal production.Wageningen Academic Publishers.2012:134-145.
[24]Jackson M K,Winslow S G,Dockery L D,et al.Investigation of an outbreak of Aleutian disease on a commercial mink ranch[J].Am J Vet Res,1996,57(12):1706-1710.
[25]关中湘,王树志,何敬芝,等.水貂阿留申病碘凝集反应与对流免疫电泳对比试验[J].吉林农业大学学报,1985,7(3):53-57.
[26]Bloom M,Kanno H,MoriS,et al.Aleutian mink disease:puzzles and paradigms[J].Infectious Agents and Disease,1994,3(6):279-301.
[27]An S H,Ingram D G.Transmission of Aleutian disease from mink with inapparent infections[J].Am J Vet Res,1978,39(2):309-313.
[28]HensonJ B,Gorham J R,Mcguire T C,et al.Pathology and pathogenesis of Aleutian disease[J].Front Biol,1976,44:175-205.
[29]Jensen T H,Christensen L S,Chriel M,et al.Implementation and validation of a sensitive PCR detection method in the eradication campaign against Aleutian mink disease virus[J].J Virol Methods,2011,171(1):81-85.
[30]Broll S,Alexandersen S.Investigation of the pathogenesis of transplacental transmission of Aleutian mink disease parvovirus in experimentally infected mink[J].J Virol,1996,70(3):1455-1466.
[2]Alex S,BloomM E,WolfinbargerJ.Evidence of restricted viral replication in adult mink infected with Aleutian disease of mink parvovirus[J]J Virol,1988,62(5):1495-1507.
[3]Hadlow W J,Race R E,Kennedy R C.Comparative pathogenicity of four strains of Aleutian disease virus for pastel and sapphire mink[J].Infect Immun,1983,41(3):1016-1023.
[4]Hansen M,Lund E.Pregnancy rate and foetal mortality in Aleutian disease virus infected mink[J].Acta Vet Scand,1988,29(2):271-272.
[5]Padgett G A,GorhamJ R,Henson J B.Epizootiologic studies of Aleutian disease.I.Transplacental transmission of the virus[J].J Infect Dis,1967,117(1):35-38.
[6]王兰坡.应用对流免疫电泳检疫水貂阿留申病的报告[J].经济动物学报,1991,2(7):19-21.
[7]Sang Y,M J,Hou Z,et al.Phylogenetic analysis of the VP2gene of Aleutian mink disease parvoviruses isolated from 2009to 2011in China[J].Virus Genes,2012,45(1):31-37.
[8]华育平,马建.阿留申病细小病毒的分离及VP2基因遗传衍生分析[J].畜牧兽医学报,2005,36(9):960-963.
[9]Henson J,GorhamJ,Leader R.A field test for Aleutian disease[J].National Fur News,1962,34:8-9.
[10]Cho H J,Greenfield J.Eradication of Aleutian disease of mink by eliminating positive counterimmunoelectrophoresis test reactors[J].J Clin Microbiol,1978,7(1):18-22.
[11]Bloom M E,OieI K L,Wolfinbarger J,et al.Evaluation of the polymerase chain reaction(pcr)as a tool for diagnosing infections with the Aleutian mink disease parvovirus[J].Scientifur,1997,21(2):141-146.
[12]Cepica A,IwamotoT.Field evaluation of CIEP and PCR detection/removal control methods of Aleutian mink disease(AD)in Canada[C]//Larsen P F,Mller S H,Clausen T,et al.Proceedings of the Xth International scientific congress in fur animal production.Wageningen Academic Publishers,2012:196-205.
[13]Jackson M K,Ellis L C,Morrey J D,et al.Progression of Aleutian disease in natural and experimentally induced infections of mink[J].Am J Vet Res,1996,57(12):1753-1758.
[14]Hadlow W J,RaceR E,Kenneedy R C.Royal pastel mink respond variously to inoculation with Aleutian disease virus of low virulence[J].J Virol,1984,50(1):38-41.
[15]Larsen A E,Porte D D.Pathogenesis of aleutian disease of mink:identification of nonpersistent infections[J].Infect Immun,1975,11(1):92-94.
[16]As S H,Depauli F J,Wright P,et al.Characteristics of inapparent Aleutian disease virus infection in mink[J].Res Vet Sci,1978,24(2):200-204.
[17]An S H,IngaramD G.Detection of inapparent Aleutian disease virus infection in mink[J].Am J Vet Res,1977,38(10):1619-1624.
[18]籍玉林,曲维江,赵元楷,等.阿留申病抗体消长规律的研究[J].畜牧兽医学报,1995,26(1):42-46.
[19]黄培堂,王嘉玺,朱厚咄译.分子克隆实验指南[M].3版,北京:科学出版社,2002,467-469.
[20]赖平安,张鹤晓,刘艳华.水貂阿留申病对流免疫电泳操作规程GB/T 6682[S]//中华人民共和国北京出入境检验检疫局,中国;中华人民共和国国家质量监督检验检疫总局,2003-08-18.
[21]J.P K.Calculation for the chi-square test:An interactive calculation tool for chi-square tests of goodness of fit and independence[E].2001,April.
[22]Li Y,HuangJ,Jia Y,et al.Genetic characterization of Aleutian mink disease viruses isolated in China[J].Virus genes,2012,45(1):24-30.
[23]Cepica A,Zargar A,Mohamed B,et al.In vivo MALDI-TOF markers for early detection of Aleutian disease(AD)among the AD virus infected mink[C]//LarsenP F,Ller S H,Clausen T,et al.Proceedings of the Xth International Scientific Congress in fur animal production.Wageningen Academic Publishers.2012:134-145.
[24]Jackson M K,Winslow S G,Dockery L D,et al.Investigation of an outbreak of Aleutian disease on a commercial mink ranch[J].Am J Vet Res,1996,57(12):1706-1710.
[25]关中湘,王树志,何敬芝,等.水貂阿留申病碘凝集反应与对流免疫电泳对比试验[J].吉林农业大学学报,1985,7(3):53-57.
[26]Bloom M,Kanno H,MoriS,et al.Aleutian mink disease:puzzles and paradigms[J].Infectious Agents and Disease,1994,3(6):279-301.
[27]An S H,Ingram D G.Transmission of Aleutian disease from mink with inapparent infections[J].Am J Vet Res,1978,39(2):309-313.
[28]HensonJ B,Gorham J R,Mcguire T C,et al.Pathology and pathogenesis of Aleutian disease[J].Front Biol,1976,44:175-205.
[29]Jensen T H,Christensen L S,Chriel M,et al.Implementation and validation of a sensitive PCR detection method in the eradication campaign against Aleutian mink disease virus[J].J Virol Methods,2011,171(1):81-85.
[30]Broll S,Alexandersen S.Investigation of the pathogenesis of transplacental transmission of Aleutian mink disease parvovirus in experimentally infected mink[J].J Virol,1996,70(3):1455-1466.