稻米中镉快速检测金标试纸条的研制
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摘要
制备一种用于快速检测稻米中重金属镉的金标试纸条。将完全抗原Cd-iEDTA-BSA作为T线,羊抗鼠二抗作为C线,包被于硝酸纤维素膜上,纳米金颗粒标记的7E8G9单克隆抗体包被于金标垫上,组装成金标试纸条。试纸条的灵敏度为0.2μg/mL、重复性良好、与Hg~(2+)有较强交叉,最高共存浓度不得高于1.0μg/mL;与Zn~(2+)有一定交叉,最高共存浓度不得高于10μg/mL;与Cu~(2+)、Fe~(2+)、Ca~(2+)、Mg~(2+)、Al~(3+)、Pb~(2+)几乎无交叉:贮存期约为1年。试纸条检测稻米中镉与GFAAS结果一致。成功制备了金标试纸条,并应用于快速检测稻米中重金属镉。
To prepare a colloidal gold- based test strip for the rapid detection cadmium in rice,synthetic antigen Cd-iEDTA-BSA and goat anti-mouse immunoglobulins were utilized in the test(T) line and control(C)line respectively.They were coated on the nitrocellulose membrane(NC).Meanwhile,the gold nanoparticles labeled monoclonal antibody 7E8G9 was coated on colloidal gold binding cushion.The results showed that the strip had a high sensitivity of 0.2 μg/mL,with good repeatability and strong cross reactivity with Hg~(2+),whose highest concentration of coexistence was not higher than 1.0 μg/mL;weak cross reactivity with Zn~(2+),whose highest concentration of coexistence was not higher than 10.0 μg/mL;and almost no cross reactivity with Cu~(2+),Fe~(2+),Ca~(2+),Mg~(2+),Al~(3+)and Pb~(2+).The storage period was about 1 year,and the detection results of the cadmium in rice by strip was consistent with GFAAS.The colloidal gold- based test strip for the rapid detection of cadmium in rice was proved to be prepared successfully.
To prepare a colloidal gold- based test strip for the rapid detection cadmium in rice,synthetic antigen Cd-iEDTA-BSA and goat anti-mouse immunoglobulins were utilized in the test(T) line and control(C)line respectively.They were coated on the nitrocellulose membrane(NC).Meanwhile,the gold nanoparticles labeled monoclonal antibody 7E8G9 was coated on colloidal gold binding cushion.The results showed that the strip had a high sensitivity of 0.2 μg/mL,with good repeatability and strong cross reactivity with Hg~(2+),whose highest concentration of coexistence was not higher than 1.0 μg/mL;weak cross reactivity with Zn~(2+),whose highest concentration of coexistence was not higher than 10.0 μg/mL;and almost no cross reactivity with Cu~(2+),Fe~(2+),Ca~(2+),Mg~(2+),Al~(3+)and Pb~(2+).The storage period was about 1 year,and the detection results of the cadmium in rice by strip was consistent with GFAAS.The colloidal gold- based test strip for the rapid detection of cadmium in rice was proved to be prepared successfully.
引文
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[18]张慧嫦,张少恩,吴忠华,等.胶体金免疫层析法快速检测盐酸克伦特罗残留[J].中国国境卫生检疫杂志,2008,31(1):39-42
[19]Blake D A,Chakrabarti P,Khosraviani M,et al.Metal binding properties of a monoclonal antibody directed toward meta-lchelate complexes[J].The Journal of Biological Chemistry,1996,271(44):27677-27685
[20]张海棠,王申峰,姜金庆,等.镉离子单克隆抗体的制备及其阻断ELISA检测方法的建立[J].中国生物制品学杂志,2011,24(2):208-212
[21]唐勇,王建华,向军俭,等.镉离子单克隆抗体的鉴定及竞争ELISA的建立[J].免疫学杂志,2009,25(2):214-218
[22]Verheijen R,Osswaid I K,Dietrich R,et al.Development of a one step strip test for the detection of(Dihydro)streptomycin residues in raw milk[J].Food and Agricultural Immunology,2000,12(1):31-40.
[2]Fallcon M,Vinas P,Osuna E,et al.Environmental exposures to lead and cadmium measured in human placenta[J].Archives of Environmental&Occupational Health,2002,57(6):598-602
[3]宫靖.镉米杀机[J].新世纪周刊,2011(6):26-33
[4]Blake D A,BlakeⅡR C,Khosraviani M,et al.Immunoassays for metal ions[J].Analytica Chimica Acta,1998,376:13-19
[5]陈笑笑,桑丽雅,李康,等.一种快速检测食用油中黄曲霉毒素B1的胶体金试剂板的研制[J].中国粮油学报,2013,28(7):99-103
[6]肖治理,余建华,雷红涛,等.盐酸克伦特罗快速检测胶体金试纸的研制[J].现代食品科技,2013,29,(8):2004-2012
[7]龚云飞,陈宗伦,奚茜,等.三聚氰胺胶体金免疫层析试纸条的研制[J].生物工程学报,2012,7:887-898
[8]Liu Xi,Xiang Junjian,Tang Yong,et al.Colloidal gold nanoparticle probe-based immunochromatographic assay for the rapid detection of chromium ions in water and serum samples[J].Analytica Chimica Acta,2012,745:99-105
[9]刘斌,唐勇,向军俭,等.纳米钛富集水样重金属镉-胶体金免疫层析法快速检测方法的建立[J].分析测试学报,2010,29(3):247-251
[10]易翠平,苏芳,陈永发,等.重金属镉单克隆抗体的制备与性质分析[J],食品科学,2013,34(21):248-253
[11]GB 2762—2005,食品中污染物限量[S]
[12]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
[13]彭剑淳,刘晓达,丁晓萍,等.可见光光谱法评价胶体金粒径及分布[J].军事医学科学院院刊,2000,24(3):211-216
[14]Cvak B,Pum D,Molinelli A,et al.Synthesis and characterization of colloidal gold particles as labels for antibodies as used in lateral flow devices[J].Analyst,2012,137(8):882-1887
[15]Mey J D.The preparation and use of gold prob.In:Polak JM,Noorden SV eds(2nded).immunocytochemistry:modern methods and application[M].Brstol:wright PSG(2nded),1986:115
[16]贺听,熊晓东,梁敬博,等.免疫检测用纳米胶体金的制备及粒径控制[J].稀有金属,2005,25(4):471-474
[17]张付贤,王兴龙.免疫胶体金技术影响因素分析[J].中国畜牧兽医.2009,36(5):199-202
[18]张慧嫦,张少恩,吴忠华,等.胶体金免疫层析法快速检测盐酸克伦特罗残留[J].中国国境卫生检疫杂志,2008,31(1):39-42
[19]Blake D A,Chakrabarti P,Khosraviani M,et al.Metal binding properties of a monoclonal antibody directed toward meta-lchelate complexes[J].The Journal of Biological Chemistry,1996,271(44):27677-27685
[20]张海棠,王申峰,姜金庆,等.镉离子单克隆抗体的制备及其阻断ELISA检测方法的建立[J].中国生物制品学杂志,2011,24(2):208-212
[21]唐勇,王建华,向军俭,等.镉离子单克隆抗体的鉴定及竞争ELISA的建立[J].免疫学杂志,2009,25(2):214-218
[22]Verheijen R,Osswaid I K,Dietrich R,et al.Development of a one step strip test for the detection of(Dihydro)streptomycin residues in raw milk[J].Food and Agricultural Immunology,2000,12(1):31-40.