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Simultaneous Detection of Thyroid-Stimulating Hormone and Free Thyroxin in Dried Spots of Human Blood by Using Phosphorescent Nanoparticles

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Abstract—

A Phosphorescence Analysis (PHOSPHAN)-based test system was developed for simultaneous detection of thyroid-stimulating hormone (TSH) and free thyroxin (fT4), the markers of thyroid function, in dried human blood samples. The immunoassay was performed at the bottom of polystyrene 96-well microplate wells covered with analyte-specific microarrays (0.35 mm in diameter). The detection system consisted of polystyrene nanoparticles (46 ± 14 nm in diameter) with incorporated long luminescent Eu(III) chelate complexes (1960 molecules/particle). Fluorescence intensity was measured in time-resolved mode (TRF). Analytical parameter values and possible limitations of this experimental system were investigated. The system allowed the simultaneous detection of high-molecular-weight and low-molecular-weight compounds in clinically relevant concentration ranges, from 2.0 to 50 pmol/L for free T4, and 0.8 to 50 µIU for TSH. The sensitivity of the analysis was 1.0 ± 0.2 pmol/L (fT4) and 0.3 ± 0.1 µIU/mL (TSH), which corresponded to the detection thresholds of 3 × 10–18 mol T4 and 0.9 × 10–9 IU TSH. The test determined correctly the TSH and T4 concentrations in standard samples used in quality assessment programs, as well as the thyroid status of the patients included in the investigation (n = 48). Results of hormone detection in dried blood spots with our experimental test system correlated well with the results of hormone detection in serum samples using commercial kits (r = 0.9730 for TSH and r = 0.8585 for fT4). The main drawback of the system was associated with high variability of measurement results (up to 20%), which limits the possibility of its application for the quantification of the hormones level in patient blood. We believe that this system can be useful for detecting thyroid disorders both in the existing programs of neonatal screening for congenital hypothyroidism and in mass screening surveys in risk groups of the adult population, for example, in ecologically unfavorable regions.

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REFERENCES

  1. Vignali, D.A., J. Immunol. Methods, 2000, vol. 243, nos. 1–2, pp. 243–255.

    Article  CAS  Google Scholar 

  2. Xu, Y.Y., Pettersson, K., Blomberg, K., Hemmila, I., Mikola, H., and Lovgren, T., Clin. Chem., 1992, vol. 38, no. 10, pp. 2038–2043.

    Article  CAS  Google Scholar 

  3. Kricka, L.J., Joos, T., and Fortina, P., Clin. Chem., 2003, vol. 49, p. 2109.

    Article  CAS  Google Scholar 

  4. Kingsmore, S.F., Nat. Rev. Drug Discov., 2006, vol. 5, pp. 310–321.

    Article  CAS  Google Scholar 

  5. Kusnezow, W., Syagailo, Y.V., Rueffer, S., et al., Mol. Cell Proteomics, 2006, pp. 1–53.

  6. Vasiliskov, A.V., Timofeev, E.N., Surzhikov, S.A., Drobyshev, A.L., and Mirzabekov, A.D., BioTechniques, 1999, vol. 27, pp. 592–600.

    Article  CAS  Google Scholar 

  7. Ekins, R.P., Clin. Chem., 1998, vol. 44, pp. 2015–2030.

    Article  CAS  Google Scholar 

  8. Lamont, J.V. and McConnell, R.I., Clin. Chem., 2005, vol. 51, pp. 1165–1176.

    Article  Google Scholar 

  9. Osin, N.S., Pomelova, V.G., Sokolov, A.S., Bychenkova, T.A., Bekman, N.I., Sharafudinova, T.Yu., et al., Vestn. Ross. Akad. Med. Nauk, 2007, no. 12, pp. 7–9.

  10. Pomelova, V.G., Korenberg, E.I., Kuznetsova, T.I., Bychenkova, T.A., Bekman, N.I., and Osin, N.S., PLoS One, 2015, vol. 10, no. 7, pp. 1–4.

    Article  Google Scholar 

  11. Osin, N.S. and Pomelova, V.G., in Frontiers in Research. National Institute of Allergy and Infectious Diseases, NIH, Georgiev, V.S., Western, K.A., and McGowan, J.J., Eds., Totowa, NJ: Humana Press, 2008, pp. 233–240.

    Google Scholar 

  12. Bekman, N.I., Laricheva, S.Yu., Bychenkova, T.A., Pomelova, V.G., and Osin, N.S., Multi-analytic phosphorescent immunochips for screening for congenital malformations in newborns, in I Mezhdunarodnaya nauchno-prakticheskaya konferentsiya “Postgenomnye metody analiza v biologii, laboratornoi i klinicheskoi meditsine” 17–19 noyabrya 2010 g. Moskva (I Int. Sci.-Pract. Conf. “Postgenomic Methods of Analysis in Biology and in Laboratory and Clinical Medicine,” November 17–19, 2010, Moscow).

  13. Bekman, N.I., Pomelova, V.G., and Osin, N.S., Klin. Lab. Diagn., 2018, no. 3, pp. 173–178.

  14. Sterling, K. and Lazarus, J.H., Annu. Rev. Physiol., 1977, vol. 39, pp. 349–371.

    Article  CAS  Google Scholar 

  15. Aoki, Y., Belin, R.M., and Clickner, R.J., Thyroid, 2007, vol. 17, pp. 1211–1223.

    Article  Google Scholar 

  16. Pomelova, V.G. and Osin, N.S., Vestn. Ross. Akad. Med. Nauk, 2007, vol. 12, pp. 10–16.

    Google Scholar 

  17. Lehmann, S., Delaby, C., Vialaret, J., Ducos, J., and Hirtz, C., Clin. Chem. Lab. Med., 2013, vol. 51, no. 10, pp. 1897–1909.

    CAS  PubMed  Google Scholar 

  18. Bellisario, R., Colinas, R.J., and Pass, K.A., Clin. Chem., 2000, vol. 46, pp. 1422–1424.

    Article  CAS  Google Scholar 

  19. Bekman, N.I., Bychenkova, T.A. Laricheva, S.Yu., Kanaeva, T.A., Smirnova, V.G., Demina, L.A., and Pomelova, V.G., Method for the simultaneous detection of thyrotropin and total thyroxine in dry blood spots, RF Patent no. 2480772, Byull. Izobret., 2013, no. 12.

  20. Pomelova, V.G., Bekman, N.I., Laricheva, S.Yu. Bychenkova, T.A., Osin, N.S., and Zlobin, V.N., Med. Ekstr. Sost., 2009, no. 3, pp. 43–50.

  21. Oppenheimer, J.H., N. Engl. J. Med., 1968, vol. 278, pp. 1153–1162.

    Article  CAS  Google Scholar 

  22. Paramonov, D.V., Kostryukova, T.S., Bychenkova, T.A., Pomelova, V.G., and Osin, N.S., Russ. J. Biorg. Chem., 2016, no. 6, pp. 1–10.

  23. Pomelova, V.G., Osin, N.S., Bychenkova, T.A., Paramonov, D.V., and Kostryukova, T.S., Russ. J. Biorg. Chem., 2017, no. 4, pp. 367–376.

  24. Danilova, N.P., Mertz, M.V., Lokshin, D.R., and Vasilov, R.G., Russ. J. Biorg. Chem., 1996, vol. 22, no. 1, pp 34–38.

    CAS  Google Scholar 

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Authors and Affiliations

  1. State Research Institute of Biological Engineering, 125424, Moscow, Russia

    N. I. Bekman, S. Yu. Laricheva & V. G. Pomelova

  2. Immunoscreen Co., 125424, Moscow, Russia

    T. A. Bychenkova & N. S. Osin

Authors
  1. N. I. Bekman
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  2. S. Yu. Laricheva
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  3. T. A. Bychenkova
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  4. V. G. Pomelova
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  5. N. S. Osin
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Corresponding author

Correspondence to N. I. Bekman.

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COMPLIANCE WITH ETHICAL STANDARDS

Patient blood samples were used for the study. Information about patients and the samples obtained from them were analyzed retrospectively, after the deletion of all information allowing identification of the results, therefore, informed consent of the patients was not required.

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The authors declare no conflict of interest.

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Translated by I. Shipounova

Abbreviations: Bi, biotin; BSA, bovine serum albumin; DBS, dried blood spots; DMSO, dimethyl sulfoxide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, water-soluble carbodiimide; fT4, free thyroxin; Ilum, intensity of the luminescence; SD, standard deviation; Str, streptavidin; TRF, time-resolved fluorescence; TSH, thyroid stimulating hormone; TSH-arrays, microarrays for the TSH detection; Т4-arrays, microarrays for the Т4 detection; ELISA, enzyme-linked immunosorbent assay; MCA, monoclonal antibodies; PHOSPHAN, phosphorescence analysis.

Corresponding author: phone: +7 (903) 180-52-85; e-mail: nibeckman@mail.ru.

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Bekman, N.I., Laricheva, S.Y., Bychenkova, T.A. et al. Simultaneous Detection of Thyroid-Stimulating Hormone and Free Thyroxin in Dried Spots of Human Blood by Using Phosphorescent Nanoparticles. Russ J Bioorg Chem 46, 199–206 (2020). https://doi.org/10.1134/S1068162020020077

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