Abstract
We describe a sensitive method for the immunochromatographic determination of aflatoxin B1. It is based on the following steps: 1) Competitive interaction between non-labeled specific primary antibodies and target antigens in a sample and in the test zone of a membrane; 2) detection of the immune complexes on the membrane by using a secondary antibodies labeled with gold nanoparticles. The method enables precise adjustment of the required quantities of specific antibodies and the colloidal (gold) marker. It was applied in a lateral flow format to the detection of aflatoxin B1 and exhibits a limit of detection (LOD) of 160 pg · mL−1 if detected visually, and of 30 pg · mL−1 via instrumental detection. This is significantly lower than the LOD of 2 ng · mL−1 achieved by conventional lateral flow analysis using the same reagents.
Similar content being viewed by others
References
O’Farrell B (2009) Evolution in Lateral Flow–Based Immunoassay Systems. (In book “Lateral Flow Immunoassay”). In: Wong R, Tse H (eds). Humana Press, pp 1–33. doi:10.1007/978-1-59745-240-3_1
Ngom B, Guo Y, Wang X, Bi D (2010) Development and application of lateral flow test strip technology for detection of infectious agents and chemical contaminants: a review. Anal Bioanal Chem 397(3):1113–1135. doi:10.1007/s00216-010-3661-4
Posthuma-Trumpie G, Korf J, van Amerongen A (2009) Lateral flow (immuno) assay: its strengths, weaknesses, opportunities and threats. A literature survey, Anal Bioanal Chem 393(2):569–582. doi:10.1007/s00216-008-2287-2
Anfossi L, Baggiani C, Giovannoli C, D’Arco G, Giraudi G (2013) Lateral-flow immunoassays for mycotoxins and phycotoxins: a review. Anal Bioanal Chem 405(2–3):467–480. doi:10.1007/s00216-012-6033-4
Wang S, Zhang C, Zhang Y (2009) Lateral Flow Colloidal Gold-Based Immunoassay for Pesticide. In: Rasooly A, Herold K (eds) Biosensors and Biodetection, vol 504. Methods in Molecular Biology™. Humana Press, pp 237–252. doi:10.1007/978-1-60327-569-9_15
Ryu Y, Jin Z, Kang M, Kim H-S (2011) Increase in the detection sensitivity of a lateral flow assay for a cardiac marker by oriented immobilization of antibody. BioChip J 5(3):193–198. doi:10.1007/s13206-011-5301-2
Chun P (2009) Colloidal Gold and Other Labels for Lateral Flow Immunoassays. In: Wong R, Tse H (eds) Lateral Flow Immunoassay. Humana Press, pp 1–19. doi:10.1007/978-1-59745-240-3_5
Rapid lateral flow test strips. Considerations for product development (2002). Millipore Corp, Bedford
Byzova NA, Zvereva EA, Zherdev AV, Dzantiev BB (2011) Immunochromatographic technique for express determination of ampicillin in milk and dairy products. Appl Biochem Microbiol 47(6):627–634. doi:10.1134/S0003683811060032
Chandler J, Gurmin T, Robinson N (2000) The place of gold in rapid tests. IVD Technol 6:37–49
Chen H, Liu F, Koh K, Lee J, Ye Z, Yin T, Sun L (2013) Sensitive detection of tuberculosis using nanoparticle-enhanced surface plasmon resonance. Microchim Acta 180(5–6):431–436. doi:10.1007/s00604-013-0943-5
Huang K-J, Li J, Liu Y-M, Cao X, Yu S, Yu M (2012) Disposable immunoassay for hepatitis B surface antigen based on a graphene paste electrode functionalized with gold nanoparticles and a Nafion-cysteine conjugate. Microchim Acta 177(3–4):419–426. doi:10.1007/s00604-012-0805-6
Li Y, Yuan R, Chai Y, Zhuo Y, Su H, Zhang Y (2014) Horseradish peroxidase-loaded nanospheres attached to hollow gold nanoparticles as signal enhancers in an ultrasensitive immunoassay for alpha-fetoprotein. Microchim Acta 181(5–6):679–685. doi:10.1007/s00604-014-1179-8
Urusov AE, Kostenko SN, Sveshnikov PG, Zherdev AV, Dzantiev BB (2011) Immunochromatographic assay for the detection of ochratoxin A. J Anal Chem 66(8):770–776. doi:10.1134/s1061934811080144
Xiong P, Gan N, Cui H, Zhou J, Cao Y, Hu F, Li T (2014) Incubation-free electrochemical immunoassay for diethylstilbestrol in milk using gold nanoparticle-antibody conjugates for signal amplification. Microchim Acta 181(3–4):453–462. doi:10.1007/s00604-013-1131-3
Anfossi L, Baggiani C, Giovannoli C, Biagioli F, D’Arco G, Giraudi G (2013) Optimization of a lateral flow immunoassay for the ultrasensitive detection of aflatoxin M1 in milk. Anal Chim Acta 772:75–80. doi:http://dx.doi.org/10.1016/j.aca.2013.02.020
Hermanson GT (2007) Chapter 24 - Preparation of Colloidal Gold-Labeled Proteins, 2nd edn, Bioconjugate Techniques. Academic, New York, pp 924–935. doi:10.1016/B978-0-12-370501-3.00024-2
Battilani P, Barbano C, Logrieco A (2008) Chapter 1 - Risk Assessment and Safety Evaluation of Mycotoxins in Fruits. In: Rivka B-G, Nachman P (eds) Mycotoxins in Fruits and Vegetables. Academic, San Diego, pp 1–26. doi:10.1016/B978-0-12-374126-4.00001-2
Sauceda-Friebe JC, Karsunke XYZ, Vazac S, Biselli S, Niessner R, Knopp D (2011) Regenerable immuno-biochip for screening ochratoxin A in green coffee extract using an automated microarray chip reader with chemiluminescence detection. Anal Chim Acta 689(2):234–242. doi:10.1016/j.aca.2011.01.030
Krska R, Molinelli A (2009) Rapid test strips for analysis of mycotoxins in food and feed. Anal Bioanal Chem 393(1):67–71. doi:10.1007/s00216-008-2424-y
Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nat (Lond), Phys Sci 241(105):20–22
Tijssen P (1985) Practice and Theory of Enzyme Immunoassays. Science, Elsevier
Sitta Sittampalam G, Smith WC, Miyakawa TW, Smith DR, McMorris C (1996) Application of experimental design techniques to optimize a competitive ELISA. J Immunol Methods 190(2):151–161. doi:10.1016/0022-1759(95)00262-6
Wang S, Quan Y, Lee N, Kennedy IR (2006) Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloidal gold immunoassay. J Agric Food Chem 54(7):2491–2495. doi:10.1021/jf0530401
Yang H, Li D, He R, Guo Q, Wang K, Zhang X, Huang P, Cui D (2010) A novel quantum dots-based point of care test for syphilis. Nanoscale Res Lett 5(5):875–881. doi:10.1007/s11671-010-9578-1
Chuanlai X, Huting W, Chifang P, Zhengyu J, Liqiang L (2006) Colloidal gold-based immunochromatographic assay for detection of diethylstilbestrol residues. Biomed Chromatogr 20(12):1390–1394. doi:10.1002/bmc.714
Daniel M-C, Astruc D (2003) Gold nanoparticles: assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology. Chem Rev 104(1):293–346. doi:10.1021/cr030698
Urusov AE, Kostenko SN, Sveshnikov PG, Zherdev AV, Dzantiev BB (2011) Ochratoxin A immunoassay with surface plasmon resonance registration: Lowering limit of detection by the use of colloidal gold immunoconjugates. Sensors Actuators B Chem 156(1):343–349. doi:10.1016/j.snb.2011.04.044
Chiao D-J, Shyu R-H, Hu C-S, Chiang H-Y, Tang S-S (2004) Colloidal gold-based immunochromatographic assay for detection of botulinum neurotoxin type B. J Chromatogr B 809(1):37–41. doi:10.1016/j.jchromb.2004.05.033
Choi DH, Lee SK, Oh YK, Bae BW, Lee SD, Kim S, Shin Y-B, Kim M-G (2010) A dual gold nanoparticle conjugate-based lateral flow assay (LFA) method for the analysis of troponin I. Biosens Bioelectron 25(8):1999–2002. doi:10.1016/j.bios.2010.01.019
Linares EM, Kubota LT, Michaelis J, Thalhammer S (2012) Enhancement of the detection limit for lateral flow immunoassays: Evaluation and comparison of bioconjugates. J Immunol Methods 375(1–2):264–270. doi:10.1016/j.jim.2011.11.003
Blazkova M, Rauch P, Fukal L (2010) Strip-based immunoassay for rapid detection of thiabendazole. Biosens Bioelectron 25(9):2122–2128. doi:10.1016/j.bios.2010.02.011
Acknowledgments
This work was supported by the Russian Foundation for Basic Research (grants 10-03-00990, 11-08-93968, 12-08-01303, 13-04-90479) and the Federal Target Program “Scientific and scientific-pedagogical personnel of the innovative Russia” for 2009–2013 (agreement 8284).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Urusov, A.E., Zherdev, A.V. & Dzantiev, B.B. Use of gold nanoparticle-labeled secondary antibodies to improve the sensitivity of an immunochromatographic assay for aflatoxin B1. Microchim Acta 181, 1939–1946 (2014). https://doi.org/10.1007/s00604-014-1288-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00604-014-1288-4