荧光免疫分析法检测食品中黄曲霉毒素的研究进展
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摘要
黄曲霉毒素是迄今发现的真菌毒素中毒性最强的一类,是公认的I类致癌物,在粮食、坚果、油脂、乳及其制品等多种食品中均有发现。因此,研究出准确、快速、便捷的检测方法对保障食品安全和人类健康具有重要意义。荧光免疫分析法具有灵敏度高、准确性好、检测速度快、操作简单等优势,适合现场高通量快速筛查,近年来被广泛地用于检测食品中的黄曲霉毒素。该文介绍了4类常用的荧光标记物(荧光素、量子点、上转换纳米粒子和稀土离子螯合物),并详细阐述了5种荧光免疫分析法(荧光免疫吸附法、荧光免疫层析法、荧光偏振免疫分析、荧光共振能量转移免疫分析和时间分辨免疫分析)的检测原理,以及上述方法应用于检测食品中黄曲霉毒素的国内外最新研究进展,为今后的研究提供借鉴。
Aflatoxins are the most toxic toxins among all well-known mycotoxins. They are recognized as a Class I carcinogen,and are widely present in grains,nuts,oils,milk and their products. Therefore,developing an accurate,rapid,and efficient detection method is of great significance for ensuring food safety and for human health. Fluorescence immunoassay has advantages of high sensitivity,good accuracy,fast detection,and simple operation. Therefore,it is very suitable for on-site high-throughput rapid screening. In recent years,it has been increasingly used for detecting aflatoxins in foods. In this review,four commonly used tracers(fluorescein,quantum dots,upconverting nanoparticles,and rare earth ion chelates) were introduced. Then,principles of five fluorescence immunoassays(fluorescence-linked immunosorbent assay,fluorescence immunochromatographic assay,fluorescence polarization immunoassay,fluorescence resonance energy transfer immunoassay,and time-resolved fluoroimmunoassay) were reviewed.Besides,the latest research progress on detecting aflatoxins in foods using these methods nationally and internationally were emphasized to provide guidance for future researches.
Aflatoxins are the most toxic toxins among all well-known mycotoxins. They are recognized as a Class I carcinogen,and are widely present in grains,nuts,oils,milk and their products. Therefore,developing an accurate,rapid,and efficient detection method is of great significance for ensuring food safety and for human health. Fluorescence immunoassay has advantages of high sensitivity,good accuracy,fast detection,and simple operation. Therefore,it is very suitable for on-site high-throughput rapid screening. In recent years,it has been increasingly used for detecting aflatoxins in foods. In this review,four commonly used tracers(fluorescein,quantum dots,upconverting nanoparticles,and rare earth ion chelates) were introduced. Then,principles of five fluorescence immunoassays(fluorescence-linked immunosorbent assay,fluorescence immunochromatographic assay,fluorescence polarization immunoassay,fluorescence resonance energy transfer immunoassay,and time-resolved fluoroimmunoassay) were reviewed.Besides,the latest research progress on detecting aflatoxins in foods using these methods nationally and internationally were emphasized to provide guidance for future researches.
引文
[1]吴丹.黄曲霉毒素在粮食和食品中的危害及防治[J].粮食加工,2007,32(3):91-94.
[2] International Agency for Research on Cancer(IARC).IARC monographs on the evaluation of carcinogenic risks to humans. vol. 56:Some naturally occuring substances:food items and constituents,heterocyclic aromatic amines and mycotoxins[M]. Lyon:World Health Organization Press,1993.
[3]张奇,李培武,陈小媚,等.黄曲霉毒素免疫检测技术研究进展[J].农产品质量与安全,2013(3):42-46.
[4]中华人民共和国卫生部GB 2761—2017,食品安全国家标准食品中真菌毒素限量[S].北京:中国标准出版社,2017.
[5] ANFOSSI L,BAGGIANI C,et al. Lateral-flow immunoassays for mycotoxins and phycotoxins:a review[J]. Analytical and Bioanalytical Chemistry,2013,405(2/3):467-480.
[6] RUCHIKA C H,JAY S,et al. Recent advances in mycotoxins eetection[J]. Biosensors and Bioelectronic,2016:S0 956-5 663(16):30 196-30 198.
[7] HORWIT W. Official methods of analysis of AOAC international[C]. Mycotoxins. Gaithersbury:AOAC International,2000.
[8]刘哲栋,徐晖.检测黄曲霉毒素的方法及其应用[J].现代食品,2017,11(22):33-34.
[9]鲁丽媛.基于核酸适体传感器的赭曲霉毒素A检测方法研究[D].郑州:河南工业大学,2017.
[10] WANG J J,LIU B H,HSU Y T,et al. Sensitive competitive direct enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip for detecting aflatoxin M1in milk[J]. Food Control,2011,22(6):964-969.
[11]王亚楠,王晓斐,王自良.食品黄曲霉毒素总量检测方法的研究与应用[J].食品与发酵工业,2018,44(1):285-290.
[12] VDOVENKO M M,LU C C,YU F Y,et al. Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin M1in milk[J]. Food Chemistry,2014,158(5):310-314.
[13] ADNYI N,LEVKOVETS I A,RODRIGUEZ-GIL S,et al. Development of immunosensor based on OWLS technique for determining aflatoxin B1and ochratoxin A[J].Biosensors&Bioelectronics,2007,22(6):797-802.
[14]张振亚,梅兴国.现代荧光免疫分析技术应用及其新发展[J].生物技术通讯,2006,17(4):677-680.
[15]朱焯炜,解希顺,陈国庆.分子荧光光谱法及其工程应用[J].物理,2008(9):56-57.
[16]汤轶伟,张宏,崔芷萌,等.荧光标记免疫层析技术在食品安全检测中的应用研究进展[J].食品工业科技,2018,39(2):314-319.
[17] ZHANG X,WEN K,WANG Z,et al. An ultra-sensitive monoclonal antibody-based fluorescent microsphere immunochromatographic test strip assay for detecting aflatoxin M1in milk[J]. Food Control,2016,60(2):588-595.
[18] WU C,HU L,XIA J,et al. Comparison of immunochromatographic assays based on fluorescent microsphere and quantum-dot submicrobead for quantitative detection of aflatoxin M1in milk.[J]. Journal of Dairy Science,2017,100(4):2 501-2 511.
[19] LIU D,HUANG Y,CHEN M,et al. Rapid detection method for aflatoxin B1in soybean sauce based on fluorescent microspheres probe[J]. Food Control,2015,50:659-662.
[20] KUANG H,ZHAO Y,MA W,et al. Recent developments in analytical applications of quantum dots[J]. Trac Trends in Analytical Chemistry,2011,30(10):1 620-1 636.
[21] TARANOVA N A,BERLINA A N,ZHERDEV A V,et al.Traffic light'immunochromatographic test based on multicolor quantum dots for the simultaneous detection of several antibiotics in milk[J]. Biosensors&Bioelectronics,2015,63(2):255-261.
[22] BELOGLAZOVA N V,SPERANSKAYA E S,WU A,et al. Novel multiplex fluorescent immunoassays based on quantum dot nanolabels for mycotoxins determination.[J]. Biosensors&Bioelectronics,2014,62(6):59-65.
[23] YANG Y,JING L,Yu X,et al. Coating aqueous quantum dots with silica via reverse microemulsion method:toward size-controllable and robust fluorescent nanoparticles[J].Chemistry of Materials,2007,19(17):4 123-4 128.
[24] REN M,XU H,HUANG X,et al. Immunochromatographic assay for ultrasensitive detection of aflatoxin B1in maize by highly luminescent quantum dot beads[J]. Acs Applied Materials&Interfaces,2014,6(16):14 215-14 222.
[25] CHUNG J W,GERELKHUU Z,JU H O,et al. Recent advances in luminescence properties of lanthanide-doped upconversion nanocrystals and applications for bioimaging,drug delivery,and optosensing[J]. Applied Spectroscopy Reviews,2016,51(7/9):678-705.
[26] WU S,DUAN N,ZHU C,et al. Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B1and ochratoxin A using upconversion nanoparticles as multicolor labels[J]. Biosensors&Bioelectronics,2011,30(1):35-42.
[27]王莉平.上转换纳米粒子的制备及其在毒素快速检测中的应用研究[D].长沙:湖南师范大学,2012.
[28] TU D,ZHENG W,LIU Y,et al. Luminescent biodetection based on lanthanide-doped inorganic nanoprobes[J].Coordination Chemistry Reviews,2014,273-274:13-29.
[29] BUENZLI J G. Lanthanide luminescence for biomedical analyses and imaging[J]. Chemical Reviews,2010,110(5):2 729-2 755.
[30] HUANG B,XIAO H,ZHANG J,et al. Dual-label time-resolved fluoroimmunoassay for simultaneous detection of aflatoxin B1and ochratoxin A[J]. Archives of Toxicology,2009,83(6):619-624.
[31]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):3 668-3 674.
[32] TANG X,ZHANG Z,LI P,et al. Sample-pretreatment-free based high sensitive determination of aflatoxin M1in raw milk using a time-resolved fluorescent competitive immunochromatographic assay[J]. Rsc Advances,2014,5(1):558-564.
[33] ZHANG Z,LI Y,LI P,et al. Monoclonal antibody-quantum dots Cd Te conjugate-based fluoroimmunoassay for the determination of aflatoxin B1in peanuts[J]. Food Chemistry,2014,146(1):314-319.
[34] BELOGLAZOVA N V,SHMELIN P S,GORYACHEVA I Y,et al. Liposomes loaded with quantum dots for ultrasensitive on-site determination of aflatoxin M1in milk products[J]. Analytical&Bioanalytical Chemistry,2013,405(24):7 795-7 802.
[35] ATANASOVA M,VASILEVA N,GODJEVARGOVA T.Determination of aflatoxin M1in milk by a magnetic nanoparticle-based fluorescent immunoassay[J]. Analytical Letters,2017,50(3):452-469.
[36]刘晓,王立平,周蕾,等.基于上转发光技术的奶粉及牛奶中黄曲霉毒素M1快速定量检测方法研究[J].军事医学,2014,38(11):850-854.
[37]王战辉,米铁军,沈建忠,等.荧光偏振免疫分析检测粮食及其制品中的真菌毒素研究进展[J].中国农业科学,2012,45(23):4 862-4 872.
[38] NASIR M S,JOLLEY M E. Development of a fluorescence polarization assay for the determination of aflatoxins in grains[J]. Journal of Agricultural&Food Chemistry,2002,50(11):3 116-3 121.
[39] SHENG Y J,EREMIN S,SERGEI T J,et al. The development of a fluorescence polarization immunoassay for aflatoxin detection[J]. Biomedical and Environmental Sciences,2014,27(2):126-129.
[40] ZHANG X,TANG Q,MI T,et al. Dual-wavelength fluorescence polarization immunoassay to increase information content per screen:Applications for simultaneous detection of total aflatoxins and family zearalenones in maize[J]. Food Control,2018,87:100-108.
[41] BELOGLAZOVA N V,EREMIN S A. Rapid screening of aflatoxin B1in beer by fluorescence polarization immunoassay[J]. Talanta,2015,142:170-175.
[42] CHEN G,SONG F,XIONG X,et al. Fluorescent nanosensors based on fluorescence resonance energy transfer(FRET)[J]. Industrial&Engineering Chemistry Research,2013,52(33):11 228-11 245.
[43] ZEKAVATI R,SAFI S,HASHEMI SJ,et al. Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1using cadmium telluride quantum dots[J]. Microchim Acta,2013,180(1314):1 217-1 223.
[44] ASWANI KUMAR Y V V,RENUKA R M,ACHUTH J,et al. Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1in contaminated food grain samples[J]. Rsc Advances,2018,8(19):10 465-10 473.
[45] SABET F S,HOSSEINI M,KHABBAZ H,et al. FRETbased aptamer biosensor for selective and sensitive detection of aflatoxin B1in peanut and rice[J]. Food Chemistry,2017,220:527-532.
[46] XU W,XIONG Y,LAI W,et al. A homogeneous immunosensor for AFB1detection based on FRET between different-sized quantum dots.[J]. Biosensors&Bioelectronics,2014,56(12):144-150.
[47] LI T,BYUN J Y,BO B K,et al. Label-free homogeneous FRET immunoassay for the detection of mycotoxins that utilizes quenching of the intrinsic fluorescence of antibodies[J]. Biosensors&Bioelectronics,2013,42(1):403-408.
[48] HAGAN A K,ZUCHNER T. Lanthanide-based time-resolved luminescence immunoassays[J]. Analytical&Bioanalytical Chemistry,2011,400(9):2 847-2 864.
[49] ZHANG Z,TANG X,WANG D,et al. Rapid on-site sensing aflatoxin B1in food and feed via a chromatographic time-resolved fluoroimmunoassay[J]. Plos One,2015,10(4):e0 123 266.
[50] TANG X Q,LI P W,ZHANG Q et al. Time-resolved fluorescence immunochromatographic assay developed using two idiotypic nanobodies for rapid,quantitative,and simultaneous detection of aflatoxin and zearalenone in maize and its products[J]. Analytical Chemistry,2017,89(21):11 520-11 528.
[2] International Agency for Research on Cancer(IARC).IARC monographs on the evaluation of carcinogenic risks to humans. vol. 56:Some naturally occuring substances:food items and constituents,heterocyclic aromatic amines and mycotoxins[M]. Lyon:World Health Organization Press,1993.
[3]张奇,李培武,陈小媚,等.黄曲霉毒素免疫检测技术研究进展[J].农产品质量与安全,2013(3):42-46.
[4]中华人民共和国卫生部GB 2761—2017,食品安全国家标准食品中真菌毒素限量[S].北京:中国标准出版社,2017.
[5] ANFOSSI L,BAGGIANI C,et al. Lateral-flow immunoassays for mycotoxins and phycotoxins:a review[J]. Analytical and Bioanalytical Chemistry,2013,405(2/3):467-480.
[6] RUCHIKA C H,JAY S,et al. Recent advances in mycotoxins eetection[J]. Biosensors and Bioelectronic,2016:S0 956-5 663(16):30 196-30 198.
[7] HORWIT W. Official methods of analysis of AOAC international[C]. Mycotoxins. Gaithersbury:AOAC International,2000.
[8]刘哲栋,徐晖.检测黄曲霉毒素的方法及其应用[J].现代食品,2017,11(22):33-34.
[9]鲁丽媛.基于核酸适体传感器的赭曲霉毒素A检测方法研究[D].郑州:河南工业大学,2017.
[10] WANG J J,LIU B H,HSU Y T,et al. Sensitive competitive direct enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip for detecting aflatoxin M1in milk[J]. Food Control,2011,22(6):964-969.
[11]王亚楠,王晓斐,王自良.食品黄曲霉毒素总量检测方法的研究与应用[J].食品与发酵工业,2018,44(1):285-290.
[12] VDOVENKO M M,LU C C,YU F Y,et al. Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin M1in milk[J]. Food Chemistry,2014,158(5):310-314.
[13] ADNYI N,LEVKOVETS I A,RODRIGUEZ-GIL S,et al. Development of immunosensor based on OWLS technique for determining aflatoxin B1and ochratoxin A[J].Biosensors&Bioelectronics,2007,22(6):797-802.
[14]张振亚,梅兴国.现代荧光免疫分析技术应用及其新发展[J].生物技术通讯,2006,17(4):677-680.
[15]朱焯炜,解希顺,陈国庆.分子荧光光谱法及其工程应用[J].物理,2008(9):56-57.
[16]汤轶伟,张宏,崔芷萌,等.荧光标记免疫层析技术在食品安全检测中的应用研究进展[J].食品工业科技,2018,39(2):314-319.
[17] ZHANG X,WEN K,WANG Z,et al. An ultra-sensitive monoclonal antibody-based fluorescent microsphere immunochromatographic test strip assay for detecting aflatoxin M1in milk[J]. Food Control,2016,60(2):588-595.
[18] WU C,HU L,XIA J,et al. Comparison of immunochromatographic assays based on fluorescent microsphere and quantum-dot submicrobead for quantitative detection of aflatoxin M1in milk.[J]. Journal of Dairy Science,2017,100(4):2 501-2 511.
[19] LIU D,HUANG Y,CHEN M,et al. Rapid detection method for aflatoxin B1in soybean sauce based on fluorescent microspheres probe[J]. Food Control,2015,50:659-662.
[20] KUANG H,ZHAO Y,MA W,et al. Recent developments in analytical applications of quantum dots[J]. Trac Trends in Analytical Chemistry,2011,30(10):1 620-1 636.
[21] TARANOVA N A,BERLINA A N,ZHERDEV A V,et al.Traffic light'immunochromatographic test based on multicolor quantum dots for the simultaneous detection of several antibiotics in milk[J]. Biosensors&Bioelectronics,2015,63(2):255-261.
[22] BELOGLAZOVA N V,SPERANSKAYA E S,WU A,et al. Novel multiplex fluorescent immunoassays based on quantum dot nanolabels for mycotoxins determination.[J]. Biosensors&Bioelectronics,2014,62(6):59-65.
[23] YANG Y,JING L,Yu X,et al. Coating aqueous quantum dots with silica via reverse microemulsion method:toward size-controllable and robust fluorescent nanoparticles[J].Chemistry of Materials,2007,19(17):4 123-4 128.
[24] REN M,XU H,HUANG X,et al. Immunochromatographic assay for ultrasensitive detection of aflatoxin B1in maize by highly luminescent quantum dot beads[J]. Acs Applied Materials&Interfaces,2014,6(16):14 215-14 222.
[25] CHUNG J W,GERELKHUU Z,JU H O,et al. Recent advances in luminescence properties of lanthanide-doped upconversion nanocrystals and applications for bioimaging,drug delivery,and optosensing[J]. Applied Spectroscopy Reviews,2016,51(7/9):678-705.
[26] WU S,DUAN N,ZHU C,et al. Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B1and ochratoxin A using upconversion nanoparticles as multicolor labels[J]. Biosensors&Bioelectronics,2011,30(1):35-42.
[27]王莉平.上转换纳米粒子的制备及其在毒素快速检测中的应用研究[D].长沙:湖南师范大学,2012.
[28] TU D,ZHENG W,LIU Y,et al. Luminescent biodetection based on lanthanide-doped inorganic nanoprobes[J].Coordination Chemistry Reviews,2014,273-274:13-29.
[29] BUENZLI J G. Lanthanide luminescence for biomedical analyses and imaging[J]. Chemical Reviews,2010,110(5):2 729-2 755.
[30] HUANG B,XIAO H,ZHANG J,et al. Dual-label time-resolved fluoroimmunoassay for simultaneous detection of aflatoxin B1and ochratoxin A[J]. Archives of Toxicology,2009,83(6):619-624.
[31]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):3 668-3 674.
[32] TANG X,ZHANG Z,LI P,et al. Sample-pretreatment-free based high sensitive determination of aflatoxin M1in raw milk using a time-resolved fluorescent competitive immunochromatographic assay[J]. Rsc Advances,2014,5(1):558-564.
[33] ZHANG Z,LI Y,LI P,et al. Monoclonal antibody-quantum dots Cd Te conjugate-based fluoroimmunoassay for the determination of aflatoxin B1in peanuts[J]. Food Chemistry,2014,146(1):314-319.
[34] BELOGLAZOVA N V,SHMELIN P S,GORYACHEVA I Y,et al. Liposomes loaded with quantum dots for ultrasensitive on-site determination of aflatoxin M1in milk products[J]. Analytical&Bioanalytical Chemistry,2013,405(24):7 795-7 802.
[35] ATANASOVA M,VASILEVA N,GODJEVARGOVA T.Determination of aflatoxin M1in milk by a magnetic nanoparticle-based fluorescent immunoassay[J]. Analytical Letters,2017,50(3):452-469.
[36]刘晓,王立平,周蕾,等.基于上转发光技术的奶粉及牛奶中黄曲霉毒素M1快速定量检测方法研究[J].军事医学,2014,38(11):850-854.
[37]王战辉,米铁军,沈建忠,等.荧光偏振免疫分析检测粮食及其制品中的真菌毒素研究进展[J].中国农业科学,2012,45(23):4 862-4 872.
[38] NASIR M S,JOLLEY M E. Development of a fluorescence polarization assay for the determination of aflatoxins in grains[J]. Journal of Agricultural&Food Chemistry,2002,50(11):3 116-3 121.
[39] SHENG Y J,EREMIN S,SERGEI T J,et al. The development of a fluorescence polarization immunoassay for aflatoxin detection[J]. Biomedical and Environmental Sciences,2014,27(2):126-129.
[40] ZHANG X,TANG Q,MI T,et al. Dual-wavelength fluorescence polarization immunoassay to increase information content per screen:Applications for simultaneous detection of total aflatoxins and family zearalenones in maize[J]. Food Control,2018,87:100-108.
[41] BELOGLAZOVA N V,EREMIN S A. Rapid screening of aflatoxin B1in beer by fluorescence polarization immunoassay[J]. Talanta,2015,142:170-175.
[42] CHEN G,SONG F,XIONG X,et al. Fluorescent nanosensors based on fluorescence resonance energy transfer(FRET)[J]. Industrial&Engineering Chemistry Research,2013,52(33):11 228-11 245.
[43] ZEKAVATI R,SAFI S,HASHEMI SJ,et al. Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1using cadmium telluride quantum dots[J]. Microchim Acta,2013,180(1314):1 217-1 223.
[44] ASWANI KUMAR Y V V,RENUKA R M,ACHUTH J,et al. Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1in contaminated food grain samples[J]. Rsc Advances,2018,8(19):10 465-10 473.
[45] SABET F S,HOSSEINI M,KHABBAZ H,et al. FRETbased aptamer biosensor for selective and sensitive detection of aflatoxin B1in peanut and rice[J]. Food Chemistry,2017,220:527-532.
[46] XU W,XIONG Y,LAI W,et al. A homogeneous immunosensor for AFB1detection based on FRET between different-sized quantum dots.[J]. Biosensors&Bioelectronics,2014,56(12):144-150.
[47] LI T,BYUN J Y,BO B K,et al. Label-free homogeneous FRET immunoassay for the detection of mycotoxins that utilizes quenching of the intrinsic fluorescence of antibodies[J]. Biosensors&Bioelectronics,2013,42(1):403-408.
[48] HAGAN A K,ZUCHNER T. Lanthanide-based time-resolved luminescence immunoassays[J]. Analytical&Bioanalytical Chemistry,2011,400(9):2 847-2 864.
[49] ZHANG Z,TANG X,WANG D,et al. Rapid on-site sensing aflatoxin B1in food and feed via a chromatographic time-resolved fluoroimmunoassay[J]. Plos One,2015,10(4):e0 123 266.
[50] TANG X Q,LI P W,ZHANG Q et al. Time-resolved fluorescence immunochromatographic assay developed using two idiotypic nanobodies for rapid,quantitative,and simultaneous detection of aflatoxin and zearalenone in maize and its products[J]. Analytical Chemistry,2017,89(21):11 520-11 528.