Abstract
A method is proposed for reducing the detection limit of enzyme-linked immunosorbent assay (ELISA) of potato virus X, based on the multiple introduction of tyramine into immune complexes and the subsequent detection of an enzyme label. During ELISA, a step by step formation of sandwich complex containing immobilized antibodies – potato virus X – horseradish peroxidase conjugate with antibodies to the virus was carried out. Peroxidase catalyzed the multiple insertion of a tyramine-biotin label into protein molecules, providing signal amplification upon the addition of the streptavidin-polyperoxidase conjugate. The conditions of the assay that ensure a high degree of amplification and a minimum background signal were established. The use of tyramine amplification made it possible to lower the detection limit by more than 30 times (from 100 to 3 ng/ml) when assayed in the buffer and extracts of potato leaves, slightly increasing its duration. Tyramine amplification is based on the use of universal reagents and can be used to reduce the detection limit of ELISA for other antigens.
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REFERENCES
Tighe, P.J., Ryder, R.R., Todd, I., and Fairclough, L.C., Proteomics Clin. Appl., 2015, vol. 9, nos. 3-4, pp. 406–422.
Asensio, L., Gonzalez, I., Garcia, T., and Martin, R., Food Control, 2008, vol. 19, no. 1, pp. 1–8.
Wild, D., The Immunoassay Handbook, Waltham: Elsevier, 2013.
Zhang, Z., Zeng, K., and Liu, J., TrAC, Trends Anal. Chem., 2017, vol. 87, pp. 49–57.
Watanabe, E., Miyake, S., and Yogo, Y., J. Agric. Food Chem., 2013, vol. 61, no. 51, pp. 12459–12472.
Vashist, S.K. and Luong, J.H.T., Handbook of Immunoassay Technologies Approaches, Performances and Applications, New York: Academic Press, 2018.
Satija, J., Punjabi, N., Mishra, D., and Mukherji, S., RSC Adv., 2016, vol. 6, no. 88, pp. 85440–85456.
Chang, L., Li, J., and Wang, L., Anal. Chim. Acta, 2016, vol. 910, pp. 12–24.
Bobrow, M.N., Harris, T.D., Shaughnessy, K.J., and Litt, G.J., J. Immunol. Methods, 1989, vol. 125, nos. 1–2, pp. 279–285.
Zhan, L., Yang, T., Li, C.M., Wu, W.B., and Huang, C.Z., Sens. Actuators, 2018, vol. 255, pp. 1291–1297.
Speel, E.J.M., Ramaekers, F.C.S., and Hopman, A.H.N., J. Histochem. Cytochem., 1997, vol. 45, no. 10, pp. 1439–1446.
Liu, P., Li, C., Zhang, R., Tang, Q., Wei, J., Lu, Y., and Shen, P., Biosens. Bioelectron., 2018, vol. 126, pp. 543–550.
Kubota, K., Microbes Environ., 2013, vol. 28, no. 1, pp. 3–12.
Meany, D.L., Hackler, L., Zhang, H., and Chan, D.W., J. Proteome Res., 2011, vol. 10, no. 3, pp. 1425–1431.
Akama, K., Shirai, K., and Suzuki, S., Anal. Chem., 2016, vol. 88, no. 14, pp. 7123–7129.
Li, X., Chen, B., He, M., Xiao, G., and Hu, B., Talanta, 2018, vol. 176, pp. 40–46.
Lucas-Garrote, B., Morais, S., and Maquieira, A., Sens. Actuators, 2017, vol. 246, pp. 1108–1115.
Byzova, N.A., Safenkova, I.V., Chirkov, S.N., Zherdev, A.V., Blintsov, A.N., Dzantiev, B.B., and Atabekov, I.G., Appl. Biochem. Microbiol., 2009, vol. 45, no. 2, pp. 204–209.
Nikitin, N., Ksenofontov, A., Trifonova, E., Arkhipenko, M., Petrova, E., Kondakova, O., Kirpichnikov, M., Atabekov, J., Dobrov, E., and Karpova, O., FEBS Lett., 2016, vol. 590, no. 10, pp. 1543–1551.
Safenkova, I., Zherdev, A., and Dzantiev, B., Anal. Bioanal. Chem., 2012, vol. 403, no. 6, pp. 1595–1605.
Hermanson, G.T., Bioconjugate Techniques, San Diego: Academic, 2013.
Hopman, A.H.N., Ramaekers, F.C.S., and Speel, E.J.M., J. Histochem. Cytochem., 1998, vol. 46, no. 6, pp. 771–777.
Bhattacharya, R., Bhattacharya, D., and Dhar, T.K., J. Immunol. Methods, 1999, vol. 227, nos. 1–2, pp. 31–39.
Herzig, G.P.D., Aydin, M., Dunigan, S., Shah, P., Jeong, K.C., Park, S.H., Ricke, S.C., and Ahn, S., J. Food Saf., 2016, vol. 36, no. 3, pp. 383–391.
Zerbini, M., Cricca, M., Gentilomi, G., Venturoli, S., Gallinella, G., and Musiani, M., Clin. Chim. Acta, 2000, vol. 302, nos. 1–2, pp. 79–87.
Yuan, L., Xu, L., and Liu, S., Anal. Chem., 2012, vol. 84, no. 24, pp. 10737–10744.
Fu, C., Jin, S., Shi, W., Oh, J., Cao, H., and Jung, Y.M., Anal. Chem., 2018, vol. 90, no. 22, pp. 13159–13162.
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This work was supported by the Russian Science Foundation (project no. 14-14-01131).
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Panferova, N.A., Panferov, V.G., Safenkova, I.V. et al. Development of Enzyme-Linked Immunosorbent Assay with Tiramine Amplification for the Detection of Potato Virus X. Appl Biochem Microbiol 55, 434–440 (2019). https://doi.org/10.1134/S0003683819040136
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DOI: https://doi.org/10.1134/S0003683819040136