不同ELISA试剂盒检测感染猪口蹄疫病毒3ABC抗体效果比较
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摘要
为更准确研究口蹄疫病毒(FMDV)感染猪非结构蛋白抗体变化规律,选取3种不同的试剂盒,分别检测接种O型口蹄疫病毒猪FMDV非结构蛋白3ABC抗体。试验数据表明,人工感染O型口蹄疫病毒猪最早在感染后第5天检测到非结构蛋白抗体,至第7天3ABC抗体全部转为阳性;3种试剂盒检测最早3ABC抗体与症状对应关系有差异,每两种商业试剂盒呈现不同程度的相关性。其中A试剂盒显示出最大的灵敏度,而B试剂盒显示最高的特异性,C试剂盒的敏感性依可疑样品的处理方式而变化,因此对3种试剂盒进行比较,发现A试剂盒更稳定,更适合于猪FMDV非结构蛋白抗体检测。
In order to study the change rule of non-structural protein antibodies in pigs inoculated with foot-and-mouth disease virus,three different kits were used in this study.The change rule of FMDV non-structural protein 3 ABC antibodies in pigs inoculated with type O foot-and-mouth disease virus were detected by ELISA.The experimental data showed that pigs inoculated with type O foot-and-mouth disease virus firstly detected positive on the 5 th day after infection,and all 3 ABC antibodies became positive on the 7 th day;non-structural protein antibody response and symptoms were different,and each two of commercial kits showed different degrees of correlation.Among them,the A kit showed the greatest sensitivity,while the B kit showed the greatest specificity,and the sensitivity of the C kit varied depending on the process of suspicious samples.Therefore,comparison the 3 kits revealed that the A kit was more stable and more suitable for detecting FMDV non-structural protein antibodies in pigs.
In order to study the change rule of non-structural protein antibodies in pigs inoculated with foot-and-mouth disease virus,three different kits were used in this study.The change rule of FMDV non-structural protein 3 ABC antibodies in pigs inoculated with type O foot-and-mouth disease virus were detected by ELISA.The experimental data showed that pigs inoculated with type O foot-and-mouth disease virus firstly detected positive on the 5 th day after infection,and all 3 ABC antibodies became positive on the 7 th day;non-structural protein antibody response and symptoms were different,and each two of commercial kits showed different degrees of correlation.Among them,the A kit showed the greatest sensitivity,while the B kit showed the greatest specificity,and the sensitivity of the C kit varied depending on the process of suspicious samples.Therefore,comparison the 3 kits revealed that the A kit was more stable and more suitable for detecting FMDV non-structural protein antibodies in pigs.
引文
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[4] Tesar M,Berger H G,Marquardt O.Serological probes for some foot-andmouth disease virus nonstructural proteins[J].Virus Gene,1989,3(1):29-44.
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[6] O'Donnell V K,Pacheco J M,Henry T M,et al.Subcellular distribution of the foot-and-mouth disease virus 3A protein in cells infected with viruses encoding wild-type and bovine-attenuated forms of 3A[J].Virology,2001,287(1):151-162.
[7] Okimura T.Effect of neurotropin(NSP)on the in vivo and in vitroantibody responses in mice[J].J Pharmacobiodyn,1986,9(8):645-650.
[8] Mackay D K,Forsyth M A,Davies P R,et al.Differentiating infection from vaccination in foot-and-mouth disease using a panel of recombinant,non-structural proteins in ELISA[J].Vaccine,1998,16(5):446-459.
[9] Lu Z,Cao Y,Guo J,et al.Development and validation of a3ABC indirect ELISA for differentiation of foot-and-mouth disease virus infected from vaccinated animals[J].Vet Microbiol,2007,125(1-2):157-169.
[10] Moonen P,Jacobs L,Crienen A,et al.Detection of carriers of foot-and-mouth disease virus among vaccinated cattle[J].Vet Microbiol,2004,103(3-4):151-160.
[11] Sorensen K J,Madsen K G,Madsen E S,et al.Differentiation of infection from vaccination in foot-and-mouth disease by the detection of antibodies to the non-structural proteins 3D,3AB and 3ABC in ELISA using antigens expressed in baculovirus[J].Arch Virol,1988,143(8):1461-1476.
[12] De Diego M,Brocchi E,Mackay D,et al.The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle[J].Arch Virol,1997,142(10):2021-2033.
[13] Bergmann I E,de Mello P A,Neitzert E,et al.Diagnosis of persistent aphthovirus infection and its differentiation from vaccination response in cattle by use of enzyme-linked immunoelec trotransfer blot analysis with bioengineered nonstructural viral antigens[J].Am J Vet Res,1993,54(6):825-831.
[14] Domingo E,Escarmis C,Baranowski E,et al.Evolution of foot-and-mouth disease virus[J].Virus Res,2003,91(1):47-63.
[15] Mason P W,Pacheco J M,Zhao Q Z,et al.Comparisons ofthe complete genomes of Asian,African and European isolates of a recent foot-and-mouth disease virus type O pandemic strain(PanAsia)[J].J Gen Viro,2003,84(6):1583-1593.
[16] Fry E E,Stuart D I,Rowlands D J.The structure of foot-andmouth disease virus[J].Curr Top Microbiol Immunol,2005,288(1):71-101.
[17] Clavijo A,Hole K,Li M,et al.Simultaneous detection of antibodies to foot-and-mouth disease nonstructural proteins3ABC,3D,3Aand 3Bby multiplexedLuminex assay to differentiate infected from vaccinated cattle[J].Vaccine,2006,24(10):1693-1704.
[18] Clavijo A,Zhou E M,Hole K,et al.Development and use of a biotinylated 3ABC recombinant protein in a solid-phase competitive ELISA for the detection of antibodies against footand-mouth disease virus[J].J Virol Methods,2004,120(2):217-227.
[2] Gosert R,E gger D,Bienz K.A cytopathic and a cell culture adapted hepatitis A virus strain differ in cell killing but not in intracellular membrane rearrangements[J].Virology,2000,266(1):157-169.
[3] Jecht M,Probst C,Gauss-Müller V.Membrane permeability induced by hepatitis A virus proteins 2Band 2BC and proteolytic processing of HAV 2BC[J].Virology,1998,252(1):218-227.
[4] Tesar M,Berger H G,Marquardt O.Serological probes for some foot-andmouth disease virus nonstructural proteins[J].Virus Gene,1989,3(1):29-44.
[5] Lubroth J,Brown F.Identification of native foot-and-mouth disease virus non-structural protein 2Cas a serological indicator to differentiate infected from vaccinated animals[J].Res Vet Sci,1995,59(1):70-78.
[6] O'Donnell V K,Pacheco J M,Henry T M,et al.Subcellular distribution of the foot-and-mouth disease virus 3A protein in cells infected with viruses encoding wild-type and bovine-attenuated forms of 3A[J].Virology,2001,287(1):151-162.
[7] Okimura T.Effect of neurotropin(NSP)on the in vivo and in vitroantibody responses in mice[J].J Pharmacobiodyn,1986,9(8):645-650.
[8] Mackay D K,Forsyth M A,Davies P R,et al.Differentiating infection from vaccination in foot-and-mouth disease using a panel of recombinant,non-structural proteins in ELISA[J].Vaccine,1998,16(5):446-459.
[9] Lu Z,Cao Y,Guo J,et al.Development and validation of a3ABC indirect ELISA for differentiation of foot-and-mouth disease virus infected from vaccinated animals[J].Vet Microbiol,2007,125(1-2):157-169.
[10] Moonen P,Jacobs L,Crienen A,et al.Detection of carriers of foot-and-mouth disease virus among vaccinated cattle[J].Vet Microbiol,2004,103(3-4):151-160.
[11] Sorensen K J,Madsen K G,Madsen E S,et al.Differentiation of infection from vaccination in foot-and-mouth disease by the detection of antibodies to the non-structural proteins 3D,3AB and 3ABC in ELISA using antigens expressed in baculovirus[J].Arch Virol,1988,143(8):1461-1476.
[12] De Diego M,Brocchi E,Mackay D,et al.The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle[J].Arch Virol,1997,142(10):2021-2033.
[13] Bergmann I E,de Mello P A,Neitzert E,et al.Diagnosis of persistent aphthovirus infection and its differentiation from vaccination response in cattle by use of enzyme-linked immunoelec trotransfer blot analysis with bioengineered nonstructural viral antigens[J].Am J Vet Res,1993,54(6):825-831.
[14] Domingo E,Escarmis C,Baranowski E,et al.Evolution of foot-and-mouth disease virus[J].Virus Res,2003,91(1):47-63.
[15] Mason P W,Pacheco J M,Zhao Q Z,et al.Comparisons ofthe complete genomes of Asian,African and European isolates of a recent foot-and-mouth disease virus type O pandemic strain(PanAsia)[J].J Gen Viro,2003,84(6):1583-1593.
[16] Fry E E,Stuart D I,Rowlands D J.The structure of foot-andmouth disease virus[J].Curr Top Microbiol Immunol,2005,288(1):71-101.
[17] Clavijo A,Hole K,Li M,et al.Simultaneous detection of antibodies to foot-and-mouth disease nonstructural proteins3ABC,3D,3Aand 3Bby multiplexedLuminex assay to differentiate infected from vaccinated cattle[J].Vaccine,2006,24(10):1693-1704.
[18] Clavijo A,Zhou E M,Hole K,et al.Development and use of a biotinylated 3ABC recombinant protein in a solid-phase competitive ELISA for the detection of antibodies against footand-mouth disease virus[J].J Virol Methods,2004,120(2):217-227.