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. 2021 Oct 21;10(21):4840.
doi: 10.3390/jcm10214840.

The Reevaluation of Thrombin Time Using a Clot Waveform Analysis

Hideo Wada  1 , Yuhuko Ichikawa  2 , Minoru Ezaki  2 , Takeshi Matsumoto  3 , Yoshiki Yamashita  4 , Katsuya Shiraki  1 , Motomu Shimaoka  5 , Hideto Shimpo  6
Affiliations

The Reevaluation of Thrombin Time Using a Clot Waveform Analysis

Hideo Wada et al. J Clin Med. .

Abstract

Object: Although thrombin burst has attracted attention as a physiological coagulation mechanism, clinical evidence from a routine assay for it is scarce. This mechanism was therefore evaluated by a clot waveform analysis (CWA) to assess the thrombin time (TT).

Material and methods: The TT with a low concentration of thrombin was evaluated using a CWA. We evaluated the CWA-TT of plasma deficient in various clotting factors, calibration plasma, platelet-poor plasma (PPP), and platelet-rich plasma (PRP) obtained from healthy volunteers, patients with thrombocytopenia, and patients with malignant disease.

Results: Although the TT-CWA of calibration plasma was able to be evaluated with 0.01 IU/mL of thrombin, that of FVIII-deficient plasma could not be evaluated. The peak time of CWA-TT was significantly longer, and the peak height significantly lower, in various deficient plasma, especially in FVIII-deficient plasma compared to calibration plasma. The second peak of the first derivative (1st DP-2) was detected in PPP from healthy volunteers, and was shorter and higher in PRP than in PPP. The 1st DP-2 was not detected in PPP from patients with thrombocytopenia, and the 1st DP-2 in PRP was significantly lower in patients with thrombocytopenia and significantly higher in patients with malignant disease than in healthy volunteers.

Conclusion: The CWA-TT became abnormal in plasma deficient in various clotting factors, and was significantly affected by platelets, suggesting that the CWA-TT may be a useful test for hemostatic abnormalities.

Keywords: CWA; platelet; thrombin burst; thrombin time.

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Conflict of interest statement

The measurements of CWA were partially supported by Instrumentation Laboratory Japan. In the other points, the authors declare no conflict of interest.

Figures

Figure 1
Figure 1
A clot waveform analysis for thrombin time. (I) Calibration plasma; (II) FVIII-deficient plasma; (a) thrombin 0.01 IU/mL; (b) thrombin 0.05 IU/mL; (c) thrombin 0.1 IU/mL; (d) thrombin 0.5 IU/mL; (e) thrombin 1.0 IU/mL; (f) thrombin 5.0 IU/mL. Navy line, fibrin formation curve; red line, 1st derivative curve (velocity); light blue, 2nd derivative curve (acceleration).
Figure 2
Figure 2
A clot waveform analysis for thrombin time. (a) Calibration plasma; (b) FII-deficient plasma; (c) FV-deficient plasma; (d) FVII-deficient plasma; (e) FVIII-deficient plasma; (f) FIX-deficient plasma; (g) FX-deficient plasma; (h) FXI-deficient plasma; (i) FXII-deficient plasma; (j) FXIII-deficient plasma; thrombin 0.5 IU/mL. Navy line, fibrin formation curve; red line, 1st derivative curve (velocity); light blue, 2nd derivative curve (acceleration).
Figure 3
Figure 3
Mixing test between calibration plasma and plasma deficient of each factor using a clot waveform analysis for thrombin time (0.5 IU/mL of thrombin). The height of fibrin formation at 100 s was plotted. The mean values of three assays are shown. The standard deviation in each assay was less than 45 mm absorbance. DP, deficient plasma.
Figure 4
Figure 4
A clot waveform analysis of thrombin time (0.5 IU/mL) in platelet-poor plasma (I) and platelet-rich plasma (II) from healthy volunteers (a), patients with thrombocytopenia (b), and patients with malignant disease (c). Navy line, fibrin formation curve; red line, 1st derivative curve (velocity); light blue, 2nd derivative curve (acceleration); red arrow shows 1st DPT-2.
See this image and copyright information in PMC

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