title: Rapid multiple immunoenzyme assay of mycotoxins.
abstract: Mycotoxins are low molecular weight fungal metabolites that pose a threat as toxic contaminants of food products, thereby necessitating their effective monitoring and control. Microplate ELISA can be used for this purpose, but this method is characteristically time consuming, with a duration extending to several hours. This report proposes a variant of the ELISA method for the detection and quantification of three mycotoxins, ochratoxin A, aflatoxin B1 and zearalenone, in the kinetic regime. The main requirement for the proposed kinetic protocol was to provide a rapid method that combined sensitivity and accuracy. The use of biotin with an extended spacer together with a streptavidin-polyperoxidase conjugate provided high signal levels, despite these interactions occurring under non-equilibrium conditions. Duration of the individual mycotoxin assays was 20 min, whereas the analysis of all three mycotoxins in parallel reached a maximum duration of 25 min. Recovery of at least 95% mycotoxins in water-organic extracts was shown. The developed assays were successfully validated using poultry processing products and corn samples spiked with known quantities of mycotoxins. The detection limits for aflatoxin B1, ochratoxin A and zearalenone in these substances were 0.24, 1.2 and 3 ng/g, respectively.
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. urusov.alexandr@gmail.com.
authors: Alexandr E
authors: AE
authors: Urusov
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. zherdev@inbi.ras.ru.
authors: Anatoly V
authors: AV
authors: Zherdev
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. alina.petrakova@gmail.com.
authors: Alina V
authors: AV
authors: Petrakova
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. elchins@gmail.com.
authors: Elchin G
authors: EG
authors: Sadykhov
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. koroleva@inbi.ras.ru.
authors: Olga V
authors: OV
authors: Koroleva
authors: A.N. Bach Institute of Biochemistry of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia. dzantiev@inbi.ras.ru.
authors: Boris B
authors: BB
authors: Dzantiev
mesh_terms: D016604:Aflatoxin B1 / Q000032:analysis*; D055910:Antibodies, Immobilized / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D057230:Limit of Detection; D009793:Ochratoxins / Q000032:analysis*; D013997:Time Factors; D015025:Zearalenone / Q000032:analysis*
chemical_list: D055910:Antibodies, Immobilized; D009793:Ochratoxins; C025589:ochratoxin A; D015025:Zearalenone; D016604:Aflatoxin B1
references: J Immunol Methods. 1996 Apr 19;190(2):151-61
references: 8621950
references: Methods Mol Biol. 2011;747:233-58
references: 21643911
references: Anal Chim Acta. 2009 Aug 11;647(2):125-36
references: 19591697
references: Prikl Biokhim Mikrobiol. 2010 May-Jun;46(3):276-90
references: 20586280
references: Anal Bioanal Chem. 2003 Jan;375(1):44-6
references: 12520437
references: J Chromatogr A. 2012 Oct 12;1259:74-85
references: 22560344
references: J Immunol Methods. 1992 Jun 24;150(1-2):5-21
references: 1613258
references: Biophys Chem. 2010 Oct;151(3):105-10
references: 20580482
references: Anal Bioanal Chem. 2009 Mar;393(5):1531-8
references: 19104778
references: J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Dec 30;942-943:88-97
references: 24231141
references: J Chromatogr A. 2012 Dec 14;1268:84-90
references: 23141710
references: Immunochemistry. 1978 Feb;15(2):71-6
references: 631867
references: Iran J Pharm Res. 2011 Fall;10(4):653-4
references: 24250401
references: Anal Biochem. 1997 Aug 15;251(1):79-88
references: 9300086
references: J Clin Endocrinol Metab. 1971 Aug;33(2):343-55
references: 5565609
references: Immunochemistry. 1978 Feb;15(2):77-82
references: 631868
references: Mol Microbiol. 1993 May;8(5):865-73
references: 8102772
references: Food Chem Toxicol. 2010 Dec;48(12):3304-7
references: 20728501
references: J Biosci Bioeng. 2007 May;103(5):427-31
references: 17609157
references: J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Nov 25;797(1-2):289-304
references: 14630156
references: Toxicol Lett. 1995 Dec;82-83:843-51
references: 8597152
references: Anal Bioanal Chem. 2013 Jan;405(2-3):467-80
references: 22543716
references: J Immunol Methods. 1997 Dec 15;210(1):93-101
references: 9502588
references: Food Chem Toxicol. 2014 Jul;69:220-30
references: 24769018
references: Int J Food Sci Nutr. 2013 Aug;64(5):632-40
references: 23369011
references: Mass Spectrom Rev. 2013 Nov-Dec;32(6):420-52
references: 23804155
references: Methods Enzymol. 1991;196:478-85
references: 2034137
references: J Chromatogr A. 2012 Sep 14;1255:145-52
references: 22424768