牛蓝舌病病毒VP7抗体胶体金试纸条检测方法的建立
|
摘要
为建立一种快速检测牛蓝舌病病毒抗体的免疫层析方法,本研究通过胶体金标记蓝舌病病毒VP7蛋白,以兔抗牛IgG作为检测线,抗蓝舌病毒VP7蛋白单克隆抗体作为质控线,采用间接法制备了胶体金免疫层析试纸条。结果显示,该胶体金试纸条在蓝舌病病毒抗体阳性血清稀释度为1∶64时仍可检出;该试纸条与牛结核病、牛布鲁氏菌病、牛病毒性腹泻、牛巴氏杆菌病、牛地方流行性白血病的阳性血清均无交叉反应;与国外商品化试剂盒相比较,该试纸条检测结果与ELISA检测试剂盒(IDEXX)的符合率为92.7%(51/55)。结果表明,本试验初步建立的牛蓝舌病病毒抗体的胶体金免疫层析检测方法具有较好的特异性和稳定性,整个检测过程可在10 min内完成,能够快速检测样品血清,适用于现场快速检测。
To establish a colloidal gold immunochromatography test strip for rapidly detecting the antibodies against bluetongue virus in bovine,the recombinant protein VP7 was used for colloidal gold marker and the goat-cow IgG was served as test line and the VP7 monoclonal antibody was served as control line.In result, even 1∶64 diluted standard bovine bluetongue disease-positive sera could be detected.The prepared colloidal gold strip showed no cross reaction with the sera with other related diseases.Compared with the foreign commercial kit,the coincidence rate between the strip and the commercial ELISA kit(IDEXX) was 92.7%(51/55).This entire test process could be completed within 10 minutes.In conclusion,the prepared colloidal gold strip showed high specificity and stability,and it might be used for the clinical detection and large-scale epidemiological survey of bluetongue virus in bovine.
To establish a colloidal gold immunochromatography test strip for rapidly detecting the antibodies against bluetongue virus in bovine,the recombinant protein VP7 was used for colloidal gold marker and the goat-cow IgG was served as test line and the VP7 monoclonal antibody was served as control line.In result, even 1∶64 diluted standard bovine bluetongue disease-positive sera could be detected.The prepared colloidal gold strip showed no cross reaction with the sera with other related diseases.Compared with the foreign commercial kit,the coincidence rate between the strip and the commercial ELISA kit(IDEXX) was 92.7%(51/55).This entire test process could be completed within 10 minutes.In conclusion,the prepared colloidal gold strip showed high specificity and stability,and it might be used for the clinical detection and large-scale epidemiological survey of bluetongue virus in bovine.
引文
[1]吕爽.蓝舌病病毒感染诱导细胞自噬的作用和机制[D].北京:中国农业科学院,2016.L譈Shuang.The effect and mechanism of cellular autophagy induction during bluetongue virus infection[D].Beijing:Chinese Academy of Agricultural Sciences,2016.(in Chinese)
[2]窦树龙,季新成,江红旗,等.2016年我国进口奶牛检疫情况分析[J].中国奶牛,2017,329(9):29-31.DOU Shu-long,JI Xin-cheng,JIANG Hong-qi,et al.Quarantine analysis of imported dairy cows in 2016[J].China Dairy Cattle,2017,339(9):29-31.(in Chinese)
[3]王贞钧.蓝舌病病毒VP7蛋白、施马伦贝格病毒N蛋白的原核表达及间接ELISA检测方法的建立[D].长春:吉林农业大学,2015.WANG Zhen-jun.Prokaryotic expression of bluetongue virus VP7 protein,Schmallenberg virus N protein and development of indirect ELISA[D].Changchun:Jilin Agricultural University,2015.(in Chinese)
[4] SAXENA V K,DEB R,SHRIVASTAVA S,et al.Functionalizing gold nanoparticles with bluetongue virus multiple peptide antigens utilizing gold-thiol interaction:a novel approach to develop pen side test[J].Research in Veterinary Science,2012,93(3):1531-1536.
[5] World Organisation for Animal Health.OIE-listed diseases,infections and infestations in force in2018[EB/OL].http://www.oie.int/animal-healthin-the-world/oie-listed-diseases-2018.
[6]李一经,李佳璇,武啸,等.25型蓝舌病病毒VP2蛋白原核表达及其单克隆抗体制备[J].东北农业大学学报,2016,47(12):65-73.LI Yi-jing,LI Jia-xuan,WU Xiao,et al.Prokaryotic expression of VP2 protein of bluetongue disease virus serotype 25 and preparation of monoclonal antibody[J].Journal of Northeast Agricultural University,2016,47(12):65-73.(in Chinese)
[7]李文超,吕茂民,杨姝,等.蓝舌病毒血清5型毒株S7基因编码区的分子克隆与序列分析[J].生物技术通讯,2007,18(3):384-387.LI Wen-chao,L譈Mao-min,YANG Shu,et al.Molecu lar cloning and sequence analysis of the S7-ORF of bluetongue virus 5[J].Letters in Biotechnolog,2007,18(3):384-387.(in Chinese)
[8]贾赟,王贞钧,孙铭英,等.蓝舌病病毒VP7蛋白的原核表达及免疫原性鉴定[J].中国畜牧兽医,2015,42(8):2000-2005.JIA Yun,WANG Zhen-jun,SUN Ming-ying,et al.Prokaryotic expression and immunogenicity identification of bluetongue virus VP7 protein[J].China Animal Husbandry&Veterinary,2015,42(8):2000-2005.(in Chinese)
[9] NAKAYAM T,ZHAO J,TAKEUCHI D,et al.Colloidal goldbased immunochromatographic strip test compromising optimised combinations of anti S.suis capsular polysaccharide polyclonal antibodies for detection of Streptococ-cussuis[J].Biosensors and Bioelectronics,2014,60(6):175-179.
[10] LI J F,ZOU M Q,CHEN Y,et al.Gold immunochromatographic strips for enhanced detection of avian influenza and Newcastle disease viruses[J].Analytica Chimica Acta,2013,782(9):54-58.
[11]樊淑华,王永立.胶体金免疫层析技术应用研究进展[J].动物医学进展,2014,35(10):99-103.FAN Shu-hua,WANG Yong-li.Progress on colloidal gold immunochromatographic assay[J].Progress in Veterinary Medicine,2014,35(10):99-103.(in Chinese)
[12]魏后军,范志宇,王芳,等.检测兔出血症病毒抗体胶体金试纸条的研制及初步应用[J].畜牧兽医学报,2018,49(3):614-619.WEI Hou-jun,FAN Zhi-yu,WANG Fang,et al.Development and preliminary application of a colloidal gold test strip for rabbit hemorrhagic disease virus antibogy[J].Acta Veterinaria et Zootechnica Sinica,2018,49(3):614-619.(in Chinese)
[13]蒋蔚,刘迎春,陈永军,等.莱克多巴胺胶体金免疫层析法快速检测试纸条的研制[J].中国动物传染病学报,2011,19(6):59-64.JIANG Wei,LIU Ying-chun,CHEN Yong-jun,et al.Development of colloidal gold immunochromatography test strips for rapid detection of ractopamine[J].Chinese Journal of Animal Infectious Diseases,2011,19(6):59-64.(in Chinese)
[14]夏启玉,李美英,杨小亮,等.免疫层析试纸条技术及其在转基因检测中的应用[J].中国生物工程杂志,2017,37(2):101-110.XIA Qi-yu,LI Mei-ying,YANG Xiao-liang,et al.Immunochromatography test strip and its applications in detection of genetically modified organisms[J].China Biotechnology,2017,37(2):101-110.(in Chinese)
[2]窦树龙,季新成,江红旗,等.2016年我国进口奶牛检疫情况分析[J].中国奶牛,2017,329(9):29-31.DOU Shu-long,JI Xin-cheng,JIANG Hong-qi,et al.Quarantine analysis of imported dairy cows in 2016[J].China Dairy Cattle,2017,339(9):29-31.(in Chinese)
[3]王贞钧.蓝舌病病毒VP7蛋白、施马伦贝格病毒N蛋白的原核表达及间接ELISA检测方法的建立[D].长春:吉林农业大学,2015.WANG Zhen-jun.Prokaryotic expression of bluetongue virus VP7 protein,Schmallenberg virus N protein and development of indirect ELISA[D].Changchun:Jilin Agricultural University,2015.(in Chinese)
[4] SAXENA V K,DEB R,SHRIVASTAVA S,et al.Functionalizing gold nanoparticles with bluetongue virus multiple peptide antigens utilizing gold-thiol interaction:a novel approach to develop pen side test[J].Research in Veterinary Science,2012,93(3):1531-1536.
[5] World Organisation for Animal Health.OIE-listed diseases,infections and infestations in force in2018[EB/OL].http://www.oie.int/animal-healthin-the-world/oie-listed-diseases-2018.
[6]李一经,李佳璇,武啸,等.25型蓝舌病病毒VP2蛋白原核表达及其单克隆抗体制备[J].东北农业大学学报,2016,47(12):65-73.LI Yi-jing,LI Jia-xuan,WU Xiao,et al.Prokaryotic expression of VP2 protein of bluetongue disease virus serotype 25 and preparation of monoclonal antibody[J].Journal of Northeast Agricultural University,2016,47(12):65-73.(in Chinese)
[7]李文超,吕茂民,杨姝,等.蓝舌病毒血清5型毒株S7基因编码区的分子克隆与序列分析[J].生物技术通讯,2007,18(3):384-387.LI Wen-chao,L譈Mao-min,YANG Shu,et al.Molecu lar cloning and sequence analysis of the S7-ORF of bluetongue virus 5[J].Letters in Biotechnolog,2007,18(3):384-387.(in Chinese)
[8]贾赟,王贞钧,孙铭英,等.蓝舌病病毒VP7蛋白的原核表达及免疫原性鉴定[J].中国畜牧兽医,2015,42(8):2000-2005.JIA Yun,WANG Zhen-jun,SUN Ming-ying,et al.Prokaryotic expression and immunogenicity identification of bluetongue virus VP7 protein[J].China Animal Husbandry&Veterinary,2015,42(8):2000-2005.(in Chinese)
[9] NAKAYAM T,ZHAO J,TAKEUCHI D,et al.Colloidal goldbased immunochromatographic strip test compromising optimised combinations of anti S.suis capsular polysaccharide polyclonal antibodies for detection of Streptococ-cussuis[J].Biosensors and Bioelectronics,2014,60(6):175-179.
[10] LI J F,ZOU M Q,CHEN Y,et al.Gold immunochromatographic strips for enhanced detection of avian influenza and Newcastle disease viruses[J].Analytica Chimica Acta,2013,782(9):54-58.
[11]樊淑华,王永立.胶体金免疫层析技术应用研究进展[J].动物医学进展,2014,35(10):99-103.FAN Shu-hua,WANG Yong-li.Progress on colloidal gold immunochromatographic assay[J].Progress in Veterinary Medicine,2014,35(10):99-103.(in Chinese)
[12]魏后军,范志宇,王芳,等.检测兔出血症病毒抗体胶体金试纸条的研制及初步应用[J].畜牧兽医学报,2018,49(3):614-619.WEI Hou-jun,FAN Zhi-yu,WANG Fang,et al.Development and preliminary application of a colloidal gold test strip for rabbit hemorrhagic disease virus antibogy[J].Acta Veterinaria et Zootechnica Sinica,2018,49(3):614-619.(in Chinese)
[13]蒋蔚,刘迎春,陈永军,等.莱克多巴胺胶体金免疫层析法快速检测试纸条的研制[J].中国动物传染病学报,2011,19(6):59-64.JIANG Wei,LIU Ying-chun,CHEN Yong-jun,et al.Development of colloidal gold immunochromatography test strips for rapid detection of ractopamine[J].Chinese Journal of Animal Infectious Diseases,2011,19(6):59-64.(in Chinese)
[14]夏启玉,李美英,杨小亮,等.免疫层析试纸条技术及其在转基因检测中的应用[J].中国生物工程杂志,2017,37(2):101-110.XIA Qi-yu,LI Mei-ying,YANG Xiao-liang,et al.Immunochromatography test strip and its applications in detection of genetically modified organisms[J].China Biotechnology,2017,37(2):101-110.(in Chinese)
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |