基于特异性生物识别分子的黄曲霉毒素快速分析方法研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.009

摘要/Abstract

摘要： 黄曲霉毒素是黄曲霉菌和寄生曲霉菌分泌的次级代谢产物,容易污染粮食作物及其制品,主要存在于发霉的粮食、豆类、坚果及与其相关食品中。黄曲霉毒素,具有极强的毒性和致癌性,尤其是黄曲霉毒素B1,是目前已发现的最强的天然致癌物质,严重威胁人类健康。目前食品中黄曲霉毒素的检测以高效液相色谱、液质联用等仪器分析方法为主,然而仪器分析方法具有设备昂贵、操作繁琐、需要专业人员等不足。近年来以特异性生物分子（抗体、重组抗体、适配体等）识别黄曲霉毒素,并结合不同的信号报告分子,建立了一系列黄曲霉毒素分析方法,检测快速、灵敏并易于操作,在食品安全领域广泛应用。着重从黄曲霉毒素的生物识别分子和信号报告分子两方面介绍目前黄曲霉毒素的快速检测手段,并对其进行比较和评价,同时对黄曲霉毒素快速检测方法的发展方向进行展望。

关键词: 黄曲霉毒素, 抗体, 适配体, 快速分析

Abstract: Aflatoxins are secondary metabolites secreted by Aspergillus flavus and Aspergillus parasiticus,easily pollute the grains and their processed products,and mainly exist in moldy grain,legumes,nuts and associated food products. Aflatoxins are extremely toxic and carcinogenic,especially aflatoxin B1 that has been found to be the strongest natural carcinogen and severely threat to human health. Current detection methods of aflatoxins in the grains are mainly instrumental analysis by combining the high performance liquid chromatography and liquid mass spectrometry,however,instrumental analysis presents the shortcomings such as instruments cost is very high,analysis process is very complex and needs long-time-trained professionals. In recent years,integrating the rapid identification of aflatoxin based on biological binders（antibody,recombinant antibody,and aptamer）and different signal reporters,a series of analysis methods for aflatoxin were established,and they are fast,flexible,and easy to operate,therefore widely applied in food safety. Here we review the rapid detection methods mainly from 2 aspects of specific biological binders for aflatoxins and signal reporters,compare and evaluate them,also prospect the development trend of rapid methods for aflatoxin.

Key words: aflatoxin, antibody, aptamer, rapid detection

曾昆,杜道林,薛永来. 基于特异性生物识别分子的黄曲霉毒素快速分析方法研究进展[J]. 生物技术通报, 2016, 32(8): 47-55.

ZENG Kun, DU Dao-lin, XUE Yong-lai. Research Advances on Rapid Detection Methods for Aflatoxin Based on Biological Binders[J]. Biotechnology Bulletin, 2016, 32(8): 47-55.

参考文献

[1] Ardic M, Karakaya Y, Atasever M, et al. Determination of aflatoxin B 1 levels in deep-red ground pepper（isot）using immunoaffinity column combined with ELISA[J]. Food and Chemical Toxicology, 2008, 46（5）：1596-1599.
[2] Detroy RW, Microbial Toxins[M]. New York：Academic Press, 1971：163-178.
[3] Han Z, Zheng Y, Luan L, et al. An ultra-high-performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in traditional Chinese medicines[J]. Analytica Chimica Acta, 2010, 664（2）：165-171.
[4] Li P, Zhang Q, Zhang W. Immunoassays for aflatoxins[J]. TrAC Trends in Analytical Chemistry, 2009, 28（9）：1115-1126.
[5] 李书国, 陈辉, 李雪梅, 等. 粮油食品中黄曲霉毒素检测方法综述[J]. 粮油食品科技, 2009, 17（2）：62-65.
[6] Cavaliere C, Foglia P, Guarino C, et al. A sensitive confirmatory method for aflatoxins in maize based on liquid chromatography/electrospray ionization tandem mass spectrometry[J]. Rapid Communications in Mass Spectrometry, 2007, 21（4）：550-556.
[7] Ventura M, Gomez A, Anaya I, et al. Determination of aflatoxins B1, G1, B2 and G2 in medicinal herbs by liquid chromatography-tandem mass spectrometry[J]. Journal of Chromatography A, 2004, 1048（1）：25-29.
[8] Van Egmond HP, Schothorst RC, Jonker MA. Regulations relating to mycotoxins in food[J]. Analytical and Bioanalytical Chemistry, 2007, 389（1）：147-157.
[9] Zhang D, Li P, Yang Y, et al. A high selective immunochromatograp-hic assay for rapid detection of aflatoxin B 1[J]. Talanta, 2011, 85（1）：736-742.
[10] Guan D, Li P, Zhang Q, et al. An ultra-sensitive monoclonal antibody-based competitive enzyme immunoassay for aflatoxin M 1 in milk and infant milk products[J]. Food Chemistry, 2011, 125（4）：1359-1364.
[11] Min WK, Kweon DH, Park K, et al. Characterisation of monoclonal antibody against aflatoxin B 1 produced in hybridoma 2C12 and its single-chain variable fragment expressed in recombinantEscherichia coli[J]. Food Chemistry, 2011, 126（3）：1316-1323.
[12] Zhang D, Li P, Zhang Q, et al. Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure[J]. Analytica Chimica Acta, 2009, 636（1）：63-69.
[13] Li P, Zhang Q, Zhang W, et al. Development of a class-specific monoclonal antibody-based ELISA for aflatoxins in peanut[J]. Food Chemistry, 2009, 115（1）：313-317.
[14] Liu JW, Lu CC, Liu BH, et al. Development of novel monoclonal antibodies-based ultrasensitive enzyme-linked immunosorbent assay and rapid immunochromatographic strip for aflatoxin B1 detection[J]. Food Control, 2016, 59：700-707.
[15] Liu BH, Hsu YT, Lu CC, et al. Detecting aflatoxin B1 in foods and feeds by using sensitive rapid enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip[J]. Food Control, 2013, 30（1）：184-189.
[16] Wang JJ, Liu BH, Hsu YT, et al. Sensitive competitive direct enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip for detecting aflatoxin M1 in milk[J]. Food Control, 2011, 22（6）：964-969.
[17] 何小鹃, 李官成. 肿瘤基因工程抗体研究进展[J]. 国外医学：肿瘤学分册, 2002, 29（4）：245-248.
[18] De Bruin R, Spelt K, Mol J, et al. Selection of high-affinity phage antibodies from phage display libraries[J]. Nature Biotechnology, 1999, 17：397-399.
[19] Moghaddam A, Løbersli I, Gebhardt K, et al. Selection and characterisation of recombinant single-chain antibodies to the hapten Aflatoxin-B1 from naive recombinant antibody libraries[J]. Journal of Immunological Methods, 2001, 254（1）：169-181.
[20] Shaw DM, Embleton MJ, Westwater C, et al. Isolation of a high affinity scFv from a monoclonal antibody recognising the oncofoetal antigen 5T4[J]. Biochimica et Biophysica Acta（BBA）-General Subjects, 2000, 1524（2）：238-246.
[21] Rangnoi K, Jaruseranee N, O’Kennedy R, et al. One-step detection of aflatoxin-B1 using scFv-alkaline phosphatase-fusion selected from human phage display antibody library[J]. Molecular Biotechnology, 2011, 49（3）：240-249.
[22] 刘蓉. 抗黄曲霉毒素 B-1 荧光重组抗体的制备[D]. 无锡：江南大学, 2010.
[23] Stoltenburg R, Reinemann C, Strehlitz B. SELEX-a（r）evolutionary method to generate high-affinity nucleic acid ligands[J]. Biomolecular Engineering, 2007, 24（4）：381-403.
[24] Malhotra S, Pandey AK, Rajput YS, et al. Selection of aptamers for aflatoxin M1 and their characterization[J]. Journal of Molecular Recognition, 2014, 27（8）：493-500.
[25] Ma X, Wang W, Chen X, et al. Selection, identification, and application of Aflatoxin B1 aptamer[J]. European Food Research and Technology, 2014, 238（6）：919-925.
[26] Fang L, Chen H, Ying X, et al. Micro-plate chemiluminescence enzyme immunoassay for aflatoxin B1 in agricultural products[J]. Talanta, 2011, 84（1）：216-222.
[27] Shim WB, Kim MJ, Mun H, et al. An aptamer-based dipstick assay for the rapid and simple detection of aflatoxin B1[J]. Biosensors and Bioelectronics, 2014, 62：288-294.
[28] Wang Y, Liu N, Ning B, et al. Simultaneous and rapid detection of six different mycotoxins using an immunochip[J]. Biosensors and Bioelectronics, 2012, 34（1）：44-50.
[29] 李军涛, 候水平, 姬泽薇, 等. 免疫荧光层析试纸条法检测食用油中的黄曲霉毒素 B-1[J]. 中国卫生检验杂志, 2015, 7：17.
[30] Zhang Z, Li Y, Li P, et al. Monoclonal antibody-quantum dots CdTe conjugate-based fluoroimmunoassay for the determination of aflatoxin B 1 in peanuts[J]. Food Chemistry, 2014, 146：314-319.
[31] 许琳, 张兆威, 李培武, 等. 量子点探针应用于粮油中黄曲霉毒素免疫检测的研究[J]. 中国油料作物学报, 2015, 37（1）：119.
[32] Xu W, Xiong Y, Lai W, et al. A homogeneous immunosensor for AFB 1 detection based on FRET between different-sized quantum dots[J]. Biosensors and Bioelectronics, 2014, 56：144-150.
[33] Wu S, Duan N, Zhu C, et al. Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B 1 and ochratoxin A using upconversion nanoparticles as multicolor labels[J]. Biosensors and Bioelectronics, 2011, 30（1）：35-42.
[34] Liu D, Huang Y, Chen M, et al. Rapid detection method for aflatoxin B 1 in soybean sauce based on fluorescent microspheres probe[J]. Food Control, 2015, 50：659-662.
[35] 张兆威, 李培武, 张奇, 等. 农产品中黄曲霉毒素的时间分辨荧光免疫层析快速检测技术研究[J]. 中国农业科学, 2014, 47（18）：3668-3674.
[36] Paniel N, Radoi A, Marty JL. Development of an electrochemical biosensor for the detection of aflatoxin M1 in milk[J]. Sensors, 2010, 10（10）：9439-9448.
[37] Parker CO, Lanyon YH, Manning M, et al. Electrochemical immunochip sensor for aflatoxin M1 detection[J]. Analytical Chemistry, 2009, 81（13）：5291-5298.
[38] Nguyen BH, Dai Tran L, Do QP, et al. Label-free detection of aflatoxin M1 with electrochemical Fe 3 O 4 /polyaniline-based aptasensor[J]. Materials Science and Engineering：C, 2013, 33（4）：2229-2234.
[39] Lin Y, Lin Y, Tang D, et al. Simple and sensitive detection of aflatoxin B 1 within five minute using a non-conventional competitive immunosensing mode[J]. Biosensors and Bioelectronics, 2015, 74：680-686.
[40] Zhang X, Li CR, Wang WC, et al. A novel electrochemical immunosensor for highly sensitive detection of aflatoxin B 1 in corn using single-walled carbon nanotubes/chitosan[J]. Food Chemistry, 2016, 192：197-202.
[41] Singh C, Srivastava S, Ali MA, et al. Carboxylated multiwalled carbon nanotubes based biosensor for aflatoxin detection[J]. Sensors and Actuators B：Chemical, 2013, 185：258-264.
[42] 孙秀兰, 汪忠云, 方银军, 等. 溶胶凝胶法固定抗体制备黄曲霉毒素免疫传感器[J]. 分析化学, 2010（2）：245-248.
[43] Li ZJ, Zhongyun W, Xiulan S, et al. A sensitive and highly stable electrochemical impedance immunosensor based on the formation of silica gel-ionic liquid biocompatible film on the glassy carbon electrode for the determination of aflatoxin B 1 in bee pollen[J]. Talanta, 2010, 80（5）：1632-1637.
[44] Yu L, Zhang Y, Hu C, et al. Highly sensitive electrochemical impedance spectroscopy immunosensor for the detection of AFB 1 in olive oil[J]. Food Chemistry, 2015, 176：22-26.
[45] Wang D, Hu W, Xiong Y, et al. Multifunctionalized reduced graphene oxide-doped polypyrrole/pyrrolepropylic acid nanocomposite impedimetric immunosensor to ultra-sensitively detect small molecular aflatoxin B 1[J]. Biosensors and Bioelectronics, 2015, 63：185-189.
[46] Castillo G, Spinella K, Poturnayová A, et al. Detection of aflatoxin B 1 by aptamer-based biosensor using PAMAM dendrimers as immobilization platform[J]. Food Control, 2015, 52：9-18.
[47] Bacher G, Pal S, Kanungo L, et al. A label-free silver wire based impedimetric immunosensor for detection of aflatoxin M1 in milk[J]. Sensors and Actuators B：Chemical, 2012, 168：223-230.
[48] Rameil S, Schubert P, Grundmann P, et al. Use of 3-（4-hydroxyp-henyl）propionic acid as electron donating compound in a potentiometric aflatoxin M 1-immunosensor[J]. Analytica Chimica Acta, 2010, 661（1）：122-127.
[49] Lv X, Li Y, Cao W, et al. A label-free electrochemiluminescence immunosensor based on silver nanoparticle hybridized mesoporous carbon for the detection of Aflatoxin B 1[J]. Sensors and Actuators B：Chemical, 2014, 202：53-59.
[50] Chauhan R, Solanki PR, Singh J, et al. A novel electrochemical piezoelectric label free immunosensor for aflatoxin B1 detection in groundnut[J]. Food Control, 2015, 52：60-70.
[51] Park JH, Kim YP, Kim IH, et al. Rapid detection of aflatoxin B 1 by a bifunctional protein crosslinker-based surface plasmon resonance biosensor[J]. Food Control, 2014, 36（1）：183-190.
[52] Jin X, Jin X, Chen L, et al. Piezoelectric immunosensor with gold nanoparticles enhanced competitive immunoreaction technique for quantification of aflatoxin B 1[J]. Biosensors and Bioelectronics, 2009, 24（8）：2580-2585.
[53] Ye Y, Zhou Y, Mo Z, et al. Rapid detection of aflatoxin B 1 on membrane by dot-immunogold filtration assay[J]. Talanta, 2010, 81（3）：792-798.
[54] 黄艳梅, 刘道峰, 赖卫华, 等. 集成免疫磁珠富集和免疫层析的黄曲霉毒素 M1 快速检测法[J]. 分析化学, 2014, 42（5）：654-659.
[55] Zhang D, Li P, Zhang Q, et al. Ultrasensitive nanogold probe-based immunochromatographic assay for simultaneous detection of total aflatoxins in peanuts[J]. Biosensors and Bioelectronics, 2011, 26（6）：2877-2882.
[56] Zhang D, Li P, Liu W, et al. Development of a detector-free semiquantitative immunochromatographic assay with major aflatoxins as target analytes[J]. Sensors and Actuators B：Chemical, 2013, 185：432-437.
[57] Xu X, Liu X, Li Y, et al. A simple and rapid optical biosensor for detection of aflatoxin B1 based on competitive dispersion of gold nanorods[J]. Biosensors and Bioelectronics, 2013, 47：361-367.
[58] Hu H, Garcia-Uribe A, Deng Y, et al. Layer-by-layer assembled smectite-polymer nanocomposite film for rapid fluorometric detection of aflatoxin B 1[J]. Sensors and Actuators B：Chemical, 2012, 166：205-211.
[59] Larou E, Yiakoumettis I, Kaltsas G, et al. High throughput cellular biosensor for the ultra-sensitive, ultra-rapid detection of aflatoxin M1[J]. Food Control, 2013, 29（1）：208-212.