猪瘟抗体检测试纸研发及制备工艺的优化
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摘要
为研制灵敏、准确、稳定的猪瘟抗体检测试纸,对胶体金标记猪瘟病毒(CSFV)Erns-E2蛋白的pH、封闭液、保存液及样品垫等试纸生产各环节的保护剂、稳定剂及缓冲系统进行优化,确定猪瘟抗体检测试纸生产的工艺参数,使试纸的准确性、灵敏度及稳定性满足批量生产要求。该试纸在室温干燥条件下可稳定保存1a以上,为实现猪瘟抗体检测试纸的商业化及广泛应用奠定基础。
Colloidal gold-labeled immunochromatographic test strip is a new immunological detection technique,which is simple,rapid,convenient,accurate,and it required no special equipment for ordinarytechnicians.For making the immunochromatographic test strips to detect classical swine fever virus(CSFV),while its specific antibodies were more sensitive,accurate and stable,we optimized the pH value,blocking liquid and preservation solutions of gold-labeled CSFV Erns-E2 protein,as well as the protective agent,stabilizers and buffer systems for each production step.The final produced strips can be stored in room temperature for more than one year.This will provide an important basis for extensive use and commercialization of the CSFV-specific antibody detection strips.
Colloidal gold-labeled immunochromatographic test strip is a new immunological detection technique,which is simple,rapid,convenient,accurate,and it required no special equipment for ordinarytechnicians.For making the immunochromatographic test strips to detect classical swine fever virus(CSFV),while its specific antibodies were more sensitive,accurate and stable,we optimized the pH value,blocking liquid and preservation solutions of gold-labeled CSFV Erns-E2 protein,as well as the protective agent,stabilizers and buffer systems for each production step.The final produced strips can be stored in room temperature for more than one year.This will provide an important basis for extensive use and commercialization of the CSFV-specific antibody detection strips.
引文
[1]于佳慧.干扰素诱导蛋白2′-5′寡腺苷酸合成酶样蛋白的抗猪瘟病毒活性研究[D].哈尔滨:中国农业科学院哈尔滨兽医研究所,2017.YU J H.Antiviralactivity of IFN-inducible porcine 2′-5′-oligoadenylate synthase-like protein against classical swine fever virus[D].Harbin:Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,2017.
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[5]LI X,WANG L,SHI X,et al.Development of an immunochromatographic strip for rapid detection of antibodies against classical swine fever virus[J].Journal of Virological Methods,2012,180(1/2):32-37.
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[8]黄印尧,张长弓,方莹,等.用于检测猪瘟抗体的免疫金标试纸条的研制与应用研究[J].福建畜牧兽医,2002,24(1):3-4.HUANG Y R,ZHANG CH G,FANG Y,et al.Development and application of immunomatographic test strip for the detection of CSFV antibody[J].Fujian Journal of Animal Husbandry and Veterinary Medicine,2002,24(1):3-4.
[9]张改平.免疫层析试纸快速检测技术[M].郑州:河南科学技术出版社,2015.ZHANG G P.Immunochromatographic Lateral Flow Strip Test[M].Zhengzhou:Henan Science and Technology Press,2015.
[10]FRENS G.Preparation gold dispersions of varying particle size controlled nucleation for the regulation of the particle size in monodisperse gold solutions[J].Nature Physical Science,1973,241:20-22.
[11]范国英,陈永耀,王顺岗,等.链霉素快速检测试纸的研制及其性能测定[J].西北农业学报,2010,19(2):40-44.FAN G Y,CHEN Y Y,WANG SH G,et al.Development and trait identification of colloidal gold-labeled strip for rapid detection of streptomycin[J].Acta Agriculturae Boreali-occidentalis Sinica,2010,19(2):40-44.
[12]GUIRE P E.Stability issues for protein-based in vitro diagnostic products[J].Ivd Technology,1999,3:50-56.
[2]OPHUIS R J,MORRISSY C J,BOYLE D B.Detection and quantitative pathogenesis study of classical swine fever virus using a real time RT-PCR assay[J].Journal of Virological Methods,2006,131(1):78-85.
[3]谢晓方.CSFV抗体间接ELISA方法的建立及初步应用[D].长沙:湖南农业大学,2016.XIE X F.Establishment and preliminary application of indirect ELISA method for CSFV antibody[D].Changsha:Hunan Agricultural University,2016.
[4]宋春美,职爱民,张静,等.莱克多巴胺荧光试纸的初步研制及性能测定[J].西北农业学报,2013,22(6):22-26.SONG CH M,ZHI A M,ZHANG J,et al.Development and trait identification of fluorescence-labeled strip for detection of ractopamine[J].Acta Agriculturae Boreali-occidentalis Sinica,2013,22(6):22-26.
[5]LI X,WANG L,SHI X,et al.Development of an immunochromatographic strip for rapid detection of antibodies against classical swine fever virus[J].Journal of Virological Methods,2012,180(1/2):32-37.
[6]REN H,ZHOU S,WEN J,et al.Development of a One-step Immunochromatographicstrip Test for Rapid Detection of Antibodies against Classic Swine Fever[M].Berlin:Springer Netherlands,2014.
[7]王建军,张元来,孙有奎.3种猪瘟抗体检测方法比较试验研究[J].国外畜牧学(猪与禽),2015,35(1):41-42.WANG J J,ZHANG Y L,SUN Y K.Comparison test on three CSF antibody detection[J].Animal Science Abroad(Pigs and Poultry),2015,35(1):41-42.
[8]黄印尧,张长弓,方莹,等.用于检测猪瘟抗体的免疫金标试纸条的研制与应用研究[J].福建畜牧兽医,2002,24(1):3-4.HUANG Y R,ZHANG CH G,FANG Y,et al.Development and application of immunomatographic test strip for the detection of CSFV antibody[J].Fujian Journal of Animal Husbandry and Veterinary Medicine,2002,24(1):3-4.
[9]张改平.免疫层析试纸快速检测技术[M].郑州:河南科学技术出版社,2015.ZHANG G P.Immunochromatographic Lateral Flow Strip Test[M].Zhengzhou:Henan Science and Technology Press,2015.
[10]FRENS G.Preparation gold dispersions of varying particle size controlled nucleation for the regulation of the particle size in monodisperse gold solutions[J].Nature Physical Science,1973,241:20-22.
[11]范国英,陈永耀,王顺岗,等.链霉素快速检测试纸的研制及其性能测定[J].西北农业学报,2010,19(2):40-44.FAN G Y,CHEN Y Y,WANG SH G,et al.Development and trait identification of colloidal gold-labeled strip for rapid detection of streptomycin[J].Acta Agriculturae Boreali-occidentalis Sinica,2010,19(2):40-44.
[12]GUIRE P E.Stability issues for protein-based in vitro diagnostic products[J].Ivd Technology,1999,3:50-56.