Development and Preliminary Application of an Indirect Competitive ELISA Kit for Detecting Deoxynivalenol

HAN Li; ZHANG Wenju; WANG Ziliang; LI Yuetao; JIANG Jinqing; LI Renfeng; FAN Guoying; ZHANG Haitang; NIU Linlin; LYU Junmei; ZHOU Ye

doi:10.3969/j.issn.1006-267x.2019.11.039

Chinese Journal of Animal Nutrition >

2019 , Vol. 31 >Issue 11: 5229 - 5237

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.11.039

Development and Preliminary Application of an Indirect Competitive ELISA Kit for Detecting Deoxynivalenol

HAN Li ,
ZHANG Wenju ,
WANG Ziliang ,
LI Yuetao ,
JIANG Jinqing ,
LI Renfeng ,
FAN Guoying ,
ZHANG Haitang ,
NIU Linlin ,
LYU Junmei ,
ZHOU Ye

Expand

1. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
2. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China

Received date: 2019-05-10

Online published: 2019-11-19

Fold

Abstract

This study was conducted to develop a self-assembled blocking indirect competitive ELISA (icELISA) kit for the detection of deoxynivalenol in feed. Based on the preparation of anti-DON monoclonal antibodies, we established the standard curve of icELISA and optimized the detection conditions, which was used to examine the performance of the kit, then the determination results were compared with the high-performance liquid chromatography (HPLC) method. The results showed that the minimum detectable concentration of the kit for DON was 1.147 ng/mL, with the detection range of 1.50 to 130.31 ng/mL, and the 50% inhibiting concentration (IC₅₀) of 9.84 ng/mL. The recovery experiment by adding standard samples was carried out for four samples of wheat, corn, pig feed and chicken feed. The recovery was in the range of 77.1% to 113.2%, and the relative standard deviation (RSD) ranged from 4.2% to 12.6%. Besides, the kit had the advantages of high specificity, good stability, long effective life and negligible sample matrix interference. Finally, twenty blind samples were determined by the kit, and the results were in good agreement with the HPLC method. These results indicate that the icELISA kit developed in this experiment has good performance to meet the relevant technical requirements, and with the characteristics of accuracy, reliability, convenience and high throughput screening to detect DON. Therefore, it is suitable for rapid screening of DON for the market.

Key words： deoxynivalenol; icELISA kit; performance measurement; preliminary application

Cite this article

HAN Li , ZHANG Wenju , WANG Ziliang , LI Yuetao , JIANG Jinqing , LI Renfeng , FAN Guoying , ZHANG Haitang , NIU Linlin , LYU Junmei , ZHOU Ye . Development and Preliminary Application of an Indirect Competitive ELISA Kit for Detecting Deoxynivalenol[J]. Chinese Journal of Animal Nutrition, 2019 , 31(11) : 5229 -5237 . DOI: 10.3969/j.issn.1006-267x.2019.11.039

References

[1] ZUO H G,ZHU J X,SHI L,et al.Development of a novel immunoaffinity column for the determination of deoxynivalenol and its acetylated derivatives in cereals[J].Food Analytical Methods,2018,11(8):2252-2260.

[2] 朱金林.呕吐毒素最新研究进展及发展趋势[J].国外畜牧学(猪与禽),2013,33(12):102-104.
[3] JIANG D F,CHEN J D,LI F H,et al.Deoxynivalenol and its acetyl derivatives in bread and biscuits in Shandong province of China[J].Food Additives and Contaminants:Part B Surveillance,2017,11(1):43-48.
[4] KONG D Z,WU X L,LI Y,et al.Ultrasensitive and eco-friendly immunoassays based monoclonal antibody for detection of deoxynivalenol in cereal and feed samples[J].Food Chemistry,2019,270:130-137.
[5] MARAGOS C M,MCCORMICK S P.Monoclonal antibodies for the mycotoxins deoxynivalenol and 3-acetyl-deoxynivalenol[J].Food and Agricultural Immunology,2000,12(3):181-192.

[6] ENNOUARI A,SANCHIS V,MARÍN S,et al.Occurrence of deoxynivalenol in durum wheat from Morocco[J].Food Control,2013,32(1):115-118.

[7] PESTKA J J,SMOLINSKI A T.Deoxynivalenol:toxicology and potential effects on humans[J].Journal of Toxicology and Environmental Health:Part B,2005,8(1):39-69.

[8] JI F,LI H,XU J H,et al.Enzyme-linked immunosorbent-assay for deoxynivalenol (DON)[J].Toxins,2011,3(12):968-978.
[9] LIAO Y X,PENG Z,CHEN L K,et al.Deoxynivalenol,gut microbiota and immunotoxicity:a potential approach?[J].Food and Chemical Toxicology,2018,112:342-354.
[10] TIMA H,BERKICS A,HANNIG Z,et al.Deoxynivalenol in wheat,maize,wheat flour and pasta:surveys in Hungary in 2008-2015[J].Food Additives & Contaminants:Part B,2018,11(1):37-42.

[11] LI M,SUN M N,HONG X,et al.Survey of deoxynivalenol contamination in agricultural products in the Chinese market using an ELISA kit[J].Toxins,2018,11(1):6.
[12] FANG S,ZHANG B,REN K W,et al.Development of a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on the monoclonal antibody for the detection of the imidaclothiz residue[J].Journal of Agricultural and Food Chemistry,2011,59(5):1594-1597.

[13] RIGHETTI L,GALAVERNA G,DALL'ASTA C.Group detection of DON and its modified forms by an ELISA kit[J].Food Additives & Contaminants:Part A,2017,34(2):248-254.
[14] ZHANG X,WANG X,SUN M J,et al.A magnetic nanoparticle based enzyme-linked immunosorbent assay for sensitive quantification of zearalenone in cereal and feed samples[J].Toxins,2015,7(10):4216-4231.

[15] DOS SANTOS J S,TAKABAYASHI C R,ONO E Y S,et al.Immunoassay based on monoclonal antibodies versus LC-MS:deoxynivalenol in wheat and flour in Southern Brazil[J].Food Additives & Contaminants:Part A,2011,28(8):1083-1090.

[16] 丁丽华,于斐,张冠军,等.脱氧雪腐镰刀菌烯醇的高灵敏化学发光磁酶免疫法检测[J].郑州大学学报(医学版),2017,52(2):134-137.
[17] LI C L,WEN K,MI T J,et al.A universal multi-wavelength fluorescence polarization immunoassay for multiplexed detection of mycotoxins in maize[J].Biosensors and Bioelectronics,2016,79:258-265.
[18] ZHANG J,GAO L,ZHOU B,et al.Simultaneous detection of deoxynivalenol and zearalenone by dual-label time-resolved fluorescence immunoassay[J].Journal of the Science of Food and Agriculture,2011,91(2):193-197.

[19] XU Y,HUANG Z B,HE Q H,et al.Development of an immunochromatographic strip test for the rapid detection of deoxynivalenol in wheat and maize[J].Food Chemistry,2010,119(2):834-839.

[20] BURMISTROVA N A,RUSANOVA T Y,YURASOV N A,et al.Multi-detection of mycotoxins by membrane based flow-through immunoassay[J].Food Control,2014,46:462-469.
[21] KADOTA T,TAKEZAWA Y,HIRANO S,et al.Rapid detection of nivalenol and deoxynivalenol in wheat using surface plasmon resonance immunoassay[J].Analytica Chimica Acta,2010,673(2):173-178.

[22] YU Q,LI H,LI C L,et al.Gold nanoparticles-based lateral flow immunoassay with silver staining for simultaneous detection of fumonisin B₁ and deoxynivalenol[J].Food Control,2015,54:347-352.
[23] QIU Y L,HE Q H,XU Y,et al.Deoxynivalenol-mimic nanobody isolated from a naive phage display nanobody library and its application in immunoassay[J].Analytica Chimica Acta,2015,887:201-208.
[24] LI Y S,SHI W M,SHEN J Z,et al.Development of a rapid competitive indirect ELISA procedure for the determination of deoxynivalenol in cereals[J].Food and Agricultural Immunology,2012,23(1):41-49.

[25] ZENG K,ZOU Y M,LIU J X,et al.Enzyme-linked immunosorbent assay for triclocarban in aquatic environments[J].Water Science and Technology,2015,72(10):1682-1691.

[26] 肖智.高特异性黄曲霉毒素B₁单克隆抗体的研制[D].硕士学位论文.北京:中国农业科学院,2010.
[27] 陈善真,李春玲,贾爱卿,等.副猪嗜血杆菌OMP5基因的克隆表达及间接ELISA检测方法的建立[J].中国农业科学,2011,44(14):3036-3044.
[28] HAYASHI Y,MATSUDA R,ITO K,et al.Detection limit estimated from slope of calibration curve:an application to competitive ELISA[J].Analytical Sciences,2005,21(2):167-169.

[29] 刘智宏,陈慧珠,赵轶先,等.鸡血清中磺胺二甲嘧啶残留的酶免疫法测定[J].中国兽医学报,1998,18(4):375-377.
[30] 国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 5009.111-2016食品安全国家标准食品中脱氧雪腐镰刀菌烯醇及其乙酰化衍生物的测定[S].北京:中国标准出版社,2017.
[31] CASALE W L,PESTKA J J,HART L P.Enzyme-linked immunosorbent assay employing monoclonal antibody specific for deoxynivalenol (vomitoxin) and several analogs[J].Journal of Agricultural and Food Chemistry,1988,36(3):663-668.

[32] LEE H M,SONG S O,CHA S H,et al.Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay[J].Journal of Veterinary Science,2013,14(2):143-150.

[33] 杨丹妮,汪海峰.小麦中呕吐毒素的ELISA和HPLC法检测[J].安徽农业科学,2013,41(3):1269-1270.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References