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Effect of fibrinogen on platelet reactivity measured by the VerifyNow P2Y12 assay

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Abstract

The VerifyNow assay is based upon the ability of activated platelets to cross-link beads coated with fibrinogen. However, fibrinogen is an abundant protein of blood, and therefore it may affect test results by competing with fibrinogen of beads for binding to platelets. To test this assumption, we assessed the influence of artificial alteration of fibrinogen level in blood samples obtained from donors (n = 9) and patients on clopidogrel therapy (n = 8) on the results of the VerifyNow P2Y12 assay. Fibrinogen level was altered by adding to blood samples 1/10 volume of fibrinogen solution (10.56 g/liter) or corresponding buffer. Relative to baseline, addition of buffer significantly increased platelet reactivity, whereas addition of fibrinogen decreased it. Analysis of the relationship between change in platelet reactivity values (dBase and dPRU) and change in fibrinogen concentration (dFg) revealed strong negative correlations: dBase =–63.3 × dFg–27.1 (r =–0.924, p < 0.0005) and dPRU =–54.4 × dFg–21.8 (r =–0.764, p < 0.0005). Thus, the results of our experiments suggest that: (i) blood fibrinogen strongly influences results of the VerifyNow P2Y12 assay, and (ii) correcting for fibrinogen effect may be needed to improve the accuracy of the test in the measuring of antiplatelet effect of clopidogrel therapy.

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Abbreviations

AU:

arbitrary units of platelet aggregation

Base:

platelet reactivity mediated by PAR-1 and -4

CVD:

cardiovas-cular diseases

Fg:

fibrinogen

GP IIb-IIIa:

glycoprotein complex binding Fg

PAR-1 and -4:

receptors activated by proteases

PRU:

platelet reactivity units mediated by P2Y12

P2Y12 :

platelet ADP receptor

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Authors and Affiliations

  1. Russian Cardiology Research and Production Complex, Ministry of Healthcare, 121552, Moscow, Russia

    A. B. Dobrovolsky, P. S. Laguta, E. V. Guskova, E. V. Titaeva, A. N. Storozhilova & E. P. Panchenko

  2. Lomonosov Moscow State University, Faculty of Mechanics and Mathematics, 119899, Moscow, Russia

    E. B. Yarovaya

Authors
  1. A. B. Dobrovolsky
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  2. P. S. Laguta
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  3. E. V. Guskova
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  4. E. B. Yarovaya
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  5. E. V. Titaeva
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  6. A. N. Storozhilova
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  7. E. P. Panchenko
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Corresponding author

Correspondence to A. B. Dobrovolsky.

Additional information

Published in Russian in Biokhimiya, 2016, Vol. 81, No. 5, pp. 595-601.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM16-011, March 27, 2016.

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Dobrovolsky, A.B., Laguta, P.S., Guskova, E.V. et al. Effect of fibrinogen on platelet reactivity measured by the VerifyNow P2Y12 assay. Biochemistry Moscow 81, 439–444 (2016). https://doi.org/10.1134/S0006297916050011

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  • DOI: https://doi.org/10.1134/S0006297916050011

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