赭曲霉毒素A和玉米赤霉烯酮-二联胶体金免疫层析试纸条的制备及应用
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摘要
【背景】真菌毒素为真菌的有毒次级代谢产物,混合污染时毒性显著增强,可对人类和动物健康造成严重伤害。制备二联胶体金免疫层析试纸条,实现对常见真菌毒素混合污染的快速监测,具有重要意义。【目的】制备赭曲霉毒素A (Ochratoxin A,OTA)和玉米赤霉烯酮(Zeralenone,ZEN)金标单克隆抗体,基于免疫层析原理,采用竞争反应模式,建立二联胶体金免疫层析检测法用于污染样品中OTA和ZEN的同时快速检测。【方法】采用柠檬酸钠还原法制备胶体金颗粒,并标记获得两种真菌毒素金标单克隆抗体,通过优化相关条件,建立稳定的二联胶体金免疫层析检测方法,用于同时检测谷物和饲料样品中的OTA和ZEN。【结果】制备的OTA和ZEN二联胶体金试纸条对OTA和ZEN的检测限分别为0.625 ng/mL和1.25 ng/mL,且与谷物和饲料中其它真菌毒素(黄曲霉毒素B1、伏马毒素B1、桔青霉毒素、展青霉毒素和呕吐毒素)均无交叉反应,人工添加试验结果准确。对天然样本检测结果表明该方法与LC-MS/MS一致性良好。【结论】本研究制备的二联胶体金试纸条可用于实际样品中OTA和ZEN的同时快速筛查。
[Background] Mycotoxins produced by different species of fungi may coexist in single cereal or feed samples, which could become highly toxic for humans and animals. In this context, a dual flow immunochromatographic assay for rapid monitoring mixed mycotoxin contamination has become a necessity. [Objective] Development of the colloidal gold dual immunoassay for simultaneous detection of ochratoxin A and zearalenone in cereal and feed samples. [Methods] Mycotoxins monoclonal antibodies were labeled with colloidal gold nanoparticles. After optimizing the materials, reagents as well as the experimental conditions, a dual flow immunochromatographic assay was developed and tested for determining the ochratoxin A and zearalenone in cereal and feed samples. [Results] The detection limit for ochratoxin A and zearalenone was 0.625 ng/mL and 1.25 ng/mL, respectively. Naturally contaminated cereal and feed samples were analyzed using both immunochromatographic assay and LC-MS/MS, and showed a good agreement between these two methods. [Conclusion] This immunoassay could detect ochratoxin A and zearalenone in cereal and feed samples rapidly and accurately, more easily and at a lower cost.
[Background] Mycotoxins produced by different species of fungi may coexist in single cereal or feed samples, which could become highly toxic for humans and animals. In this context, a dual flow immunochromatographic assay for rapid monitoring mixed mycotoxin contamination has become a necessity. [Objective] Development of the colloidal gold dual immunoassay for simultaneous detection of ochratoxin A and zearalenone in cereal and feed samples. [Methods] Mycotoxins monoclonal antibodies were labeled with colloidal gold nanoparticles. After optimizing the materials, reagents as well as the experimental conditions, a dual flow immunochromatographic assay was developed and tested for determining the ochratoxin A and zearalenone in cereal and feed samples. [Results] The detection limit for ochratoxin A and zearalenone was 0.625 ng/mL and 1.25 ng/mL, respectively. Naturally contaminated cereal and feed samples were analyzed using both immunochromatographic assay and LC-MS/MS, and showed a good agreement between these two methods. [Conclusion] This immunoassay could detect ochratoxin A and zearalenone in cereal and feed samples rapidly and accurately, more easily and at a lower cost.
引文
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[13]Molinelli A,Grossalber K,Krska R.A rapid lateral flow test for the determination of total type B fumonisins in maize[J].Analytical and Bioanalytical Chemistry,2009,395(5):1309-1316
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[16]Kolosova AY,de Saeger S,Sibanda L,et al.Development of a colloidal gold-based lateral-flow immunoassay for the rapid simultaneous detection of zearalenone and deoxynivalenol[J].Analytical and Bioanalytical Chemistry,2007,389(7/8):2103-2107
[17]Shim WB,Dzantiev BB,Eremin SA,et al.One-step simultaneous immunochromatographic strip test for multianalysis of ochratoxin a and zearalenone[J].Journal of Microbiology and Biotechnology,2009,19(1):83-92
[18]Guo YR,Liu SY,Gui WJ,et al.Gold immunochromatographic assay for simultaneous detection of carbofuran and triazophos in water samples[J].Analytical Biochemistry,2009,389(1):32-39
[19]Byzova NA,Zvereva EA,Zherdev AV,et al.Rapid pretreatment-free immunochromatographic assay of chloramphenicol in milk[J].Talanta,2010,81(3):843-848
[20]Song CM,Liu QT,Zhi AM,et al.Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of olaquindox residues[J].Journal of Agricultural and Food Chemistry,2011,59(17):9319-9326
[21]Wang S,Quan Y,Lee NJ,et al.Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloidal gold immunoassay[J].Journal of Agricultural and Food Chemistry,2006,54(7):2491-2495
[22]Lattanzio VMT,Nivarlet N,Lippolis V,et al.Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals[J].Analytica Chimica Acta,2012,718:99-108
[23]Liu Y,Wang PY,Liu XP,et al.Review of current Chinese regulations of maximum residue levels and testing standard of mycotoxin in foods and feeds[J].China Brewing,2014,33(7):10-19(in Chinese)刘萤,王珮珮,刘雪平,等.我国现行食品与饲料中真菌毒素限量及检测标准概述[J].中国酿造,2014,33(7):10-19
[24]Wang YK,Shi YB,Zou Q,et al.Development of a rapid and simultaneous immunochromatographic assay for the determination of zearalenone and fumonisin B1 in corn,wheat and feedstuff samples[J].Food Control,2013,31(1):180-188
[2]Amézqueta S,González-Pe?as E,Murillo-Arbizu M,et al.Ochratoxin a decontamination:a review[J].Food Control,2009,20(4):326-333
[3]He L,Zheng W,Liu YF,et al.Progress in ochratoxins A[J].China Herbivore Science,2017,37(5):59-61,69(in Chinese)贺亮,郑文,刘玉峰,等.赭曲霉毒素A研究进展[J].中国草食动物科学,2017,37(5):59-61,69
[4]Collins TFX,Sprando RL,Black TN,et al.Effects of zearalenone on in utero development in rats[J].Food and Chemical Toxicology,2006,44(9):1455-1465
[5]Yang LJ,Huang LB,Li SM,et al.Effects of zearalenone on ovary index,distribution and expression of progesterone receptors in ovaries of weaned gilts[J].Chinese Journal of Animal Nutrition,2017,29(12):4510-4517(in Chinese)杨立杰,黄丽波,李思梦,等.玉米赤霉烯酮对断奶小母猪卵巢指数及孕激素受体分布和表达的影响[J].动物营养学报,2017,29(12):4510-4517
[6]Smith MC,Madec S,Coton E,et al.Natural co-occurrence of mycotoxins in foods and feeds and their in vitro combined toxicological effects[J].Toxins,2016,8(4):94
[7]Zhang J,Gao L,Zhou B,et al.Study on rapidly simultaneous detection of deoxynivalenol and Zearalenone[J].China Biotechnology,2009,29(11):82-88(in Chinese)张珏,高雷,周彬,等.快速同时检测玉米赤霉烯酮和脱氧雪腐镰刀菌烯醇的研究[J].中国生物工程杂志,2009,29(11):82-88
[8]Calbiani F,Careri M,Elviri L,et al.Development and in-house validation of a liquid chromatography-electrospray-tandem mass spectrometry method for the simultaneous determination of Sudan I,Sudan II,Sudan III and Sudan IV in hot chilli products[J].Journal of Chromatography A,2004,1042(1/2):123-130
[9]Vaclavik L,Vaclavikova M,Begley TH,et al.Determination of multiple mycotoxins in dietary supplements containing green coffee bean extracts using ultrahigh-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)[J].Journal of Agricultural and Food Chemistry,2013,61(20):4822-4830
[10]Li RJ,Zhang XY,Yang C,et al.Development of a colloidal gold-based immunochromatographic assay for the rapid detection of Edwardsiella ictaluri[J].Journal of Nanoscience and Nanotechnology,2018,18(6):3800-3805
[11]Cao DK,Su JZ,Zhang Y,et al.Research progress in colloidal gold immune chromatographic techniques in food safety detection[J].Journal of Preventive Medicine of Chinese People’s Liberation Army,2017,35(10):1322-1325(in Chinese)曹德康,苏建忠,张瑛,等.胶体金免疫层析技术在食品安全检测中的应用研究进展[J].解放军预防医学杂志,2017,35(10):1322-1325
[12]Zhang DH,Li PW,Yang Y,et al.A high selective immunochromatographic assay for rapid detection of aflatoxin B1[J].Talanta,2011,85(1):736-742
[13]Molinelli A,Grossalber K,Krska R.A rapid lateral flow test for the determination of total type B fumonisins in maize[J].Analytical and Bioanalytical Chemistry,2009,395(5):1309-1316
[14]Luo SJ,Chen FR,Liu CM,et al.Effect of colloidal gold particle size on the performance of chloramphenicol test strips[J].Food Science,2011,32(12):120-124(in Chinese)罗舜菁,陈发荣,刘成梅,等.胶体金粒径对氯霉素胶体金试纸条性能的影响[J].食品科学,2011,32(12):120-124
[15]Shim WB,Kim KY,Chung DH.Development and validation of a gold nanoparticle immunochromatographic assay(ICG)for the detection of zearalenone[J].Journal of Agricultural and Food Chemistry,2009,57(10):4035-4041
[16]Kolosova AY,de Saeger S,Sibanda L,et al.Development of a colloidal gold-based lateral-flow immunoassay for the rapid simultaneous detection of zearalenone and deoxynivalenol[J].Analytical and Bioanalytical Chemistry,2007,389(7/8):2103-2107
[17]Shim WB,Dzantiev BB,Eremin SA,et al.One-step simultaneous immunochromatographic strip test for multianalysis of ochratoxin a and zearalenone[J].Journal of Microbiology and Biotechnology,2009,19(1):83-92
[18]Guo YR,Liu SY,Gui WJ,et al.Gold immunochromatographic assay for simultaneous detection of carbofuran and triazophos in water samples[J].Analytical Biochemistry,2009,389(1):32-39
[19]Byzova NA,Zvereva EA,Zherdev AV,et al.Rapid pretreatment-free immunochromatographic assay of chloramphenicol in milk[J].Talanta,2010,81(3):843-848
[20]Song CM,Liu QT,Zhi AM,et al.Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of olaquindox residues[J].Journal of Agricultural and Food Chemistry,2011,59(17):9319-9326
[21]Wang S,Quan Y,Lee NJ,et al.Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloidal gold immunoassay[J].Journal of Agricultural and Food Chemistry,2006,54(7):2491-2495
[22]Lattanzio VMT,Nivarlet N,Lippolis V,et al.Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals[J].Analytica Chimica Acta,2012,718:99-108
[23]Liu Y,Wang PY,Liu XP,et al.Review of current Chinese regulations of maximum residue levels and testing standard of mycotoxin in foods and feeds[J].China Brewing,2014,33(7):10-19(in Chinese)刘萤,王珮珮,刘雪平,等.我国现行食品与饲料中真菌毒素限量及检测标准概述[J].中国酿造,2014,33(7):10-19
[24]Wang YK,Shi YB,Zou Q,et al.Development of a rapid and simultaneous immunochromatographic assay for the determination of zearalenone and fumonisin B1 in corn,wheat and feedstuff samples[J].Food Control,2013,31(1):180-188