Cn 6000

Blood samples of 6.4 mL were collected via a radial artery cannula system with a closed circuit for blood sampling during CPB and were collected to obtain results at the same time points for fibrinogen concentration measurements for Fibcare, fibrinogen concentration measurements for our hospital’s central laboratory, and ActF-MA. Fibcare was used to measure fibrinogen concentration, and ActF-MA was used to predict the fibrinogen concentration in the operating room. Traditional laboratory-based testing was used to measure fibrinogen concentration; specimens were submitted to our hospital’s central laboratory and measured by the laboratory technician using Clauss fibrinogen assay (Analyzer: CN-6000; measuring reagent: Thrombocheck Fib [L], Sysmex Corporation, Kobe, Japan). Fibcare has a lower and upper limit of 80 and 300, respectively. If the result was <80, a value of 40 was imputed, while if the result exceeded the upper limit, the result was excluded from the analysis. ActF-MA by TEG6s has a lower limit of 2.0. If the result was <2.0, a value of 1.0 was imputed.

Conventional coagulation tests were performed at the Central Laboratory of CNUH using automated complete blood count analyzers XN-1000 (Sysmex, Kobe, Japan), XE-2100 (Sysmex), DxH-800 (Beckman Coulter, Brea, CA, USA), or DxH-900 (Beckman Coulter) for Hb, Hct, and platelet counts; Platelet Function Analyzer (PFA)-100 (Siemens, Deerfield, IL, USA) for platelet function tests with collagen/epinephrine; and automated coagulation analyzers CS-5100 (Sysmex) or CN-6000 (Sysmex) for prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen, fibrin/fibrinogen degradation products (FDPs), D-dimer, and antithrombin III.
ROTEM was performed at the discretion of the clinician, after massive transfusion, as a point-of-care test at the Trauma Intensive Care Unit of CNUH. Clot amplitudes after 5 (A5), 10 (A10), and 15 (A15) mins of extrinsically activated ROTEM (EXTEM) and fibrin-specific ROTEM (FIBTEM) were collected. The maximum clot firmness (MCF) after EXTEM and FIBTEM was also reviewed. Platelet-specific ROTEM (PLTEM) was calculated by subtracting FIBTEM from EXTEM [27 (link)].

Coagulation assays for APTT (Thrombocheck APTT-SLA using ellagic acid and silica as triggers; Sysmex, Kobe, Japan), FVIII OSA (FVIII-deficient plasma; SIEMENS, Marburg, Germany), and FVIII chromogenic assay (CSA) (Revohem FVIII chromogenic; Sysmex, Kobe, Japan) using human FX and FIXa were performed using commercially available diagnostic kits according to instructions provided by the manufacturers. Additionally, we used the parameters Adjusted |min1| (Ad|mn1|) and Adjusted |min2| (Ad|min2|), which are obtained from the APTT clotting wave form calculated by the blood coagulation analyzer CN-6000 (Sysmex, Kobe, Japan). With the modified clot waveform analysis (CWA), the minimum transmittance (0%) was set in the immediate post coagulation phase, and Ad|min1| and Ad|min2| were defined accordingly as the maximum coagulation velocity and acceleration, respectively, obtained from the first or second derivative of this adjusted clot waveform.^{14 (link)}
Samples from patients with HA with emicizumab treatment were analyzed with or without anti-idiotype antibodies (both rcAQ8 and rcAJ540).^{13 (link)}
Plasma and antibodies were mixed at a ratio of 20:1. These anti-idiotype antibodies were kindly provided by Chugai Pharmaceutical, Tokyo.