Hemosil aptt sp

Manufactured by Werfen

Sourced in United States

The Hemosil APTT-SP is a laboratory equipment product designed to measure the activated partial thromboplastin time (APTT) in blood samples. It is used to assess the intrinsic and common coagulation pathways. The product provides a quantitative analysis of the APTT measurement.

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9 protocols using hemosil aptt sp

Evaluation of the Q Smart Coagulation Analyzer

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The Q Smart system (Diagnostic Grifols, Barcelona, Spain) comprises the Q Smart analyzer and the following DG reagents (Diagnostic Grifols) for each test: DG-PT RecombiLIQ for prothrombin time (PT), DG-APTT Synth for activated partial thromboplastin time (APTT), DG-TT L Human for thrombin time (TT), DG-APTT Synth/DG-FVIII for FVIII, DG-Latex DDimer for d-dimer, and DG-Chrom AT L for Antithrombin(AT). ACL TOP 500 analyzer and Hemosil reagents from Instrumentation Laboratory (Bedford, Massachusetts) were used for comparison in the following tests: Hemosil Recombiplastin 2G for PT, Hemosil APTT-SP (liquid) for APTT, Hemosil Thrombin Time for TT, and Hemosil APTT-SP (liquid)/Hemosil FVIII Deficient Plasma for FVIII. For comparison in the d-dimer and AT tests, the BCS XP coagulometer and reagents were used (Siemens AG, Munich, Germany). Additionally, the PT assay was also performed with the CoaguChek XS point-of-care_POC (Roche, Indianapolis, Indiana).

Montalvão S.A., Francisco A.P., da Silva B.L., Huber S.C., Aguiari H.J., Fernandes M.C., Elidio P.S., Martinelli B.D., Tony I.M., Colella M.P, & Annichinno-Bizzachi J.M. (2020). From Hemophilia to Deep Venous Thrombosis Patient Samples: How to Perform an Easy Coagulometer Validation Process According to Available Guidelines. Clinical and Applied Thrombosis/Hemostasis, 26, 1076029620915512.

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Activated Partial Thromboplastin Time Kinetics

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Kinetics of turbidity development during activated partial thromboplastin time (aPTT) reactions allowed indirect assessment of clot structure. Plasma samples from individual HemA donors (HRF, Inc, Raleigh, NC, USA) were spiked with FVIII (0%–100%), and the aPTT and fibrinogen assays were performed with the HemosIL APTT-SP and Fibrinogen-C kits, respectively (Instrumentation Laboratory [IL], Bedford, MA, USA), using on-board ACL TOP tests. HemA and control clots were prepared using similar conditions as the aPTT assay using HemA and pooled plasma from healthy donors (HRF, Inc), respectively. CT was determined from the second derivative of the turbidity change over time, and turbidity waveform data were extracted from the ACL TOP using proprietary software developed by Bayer Business Services (Leverkusen, Germany). Changes in turbidity parameters were correlated with changes in CTs and with fibrinogen levels.
Direct visualization of plasma clots were achieved by confocal microscopy of plasma. HemA plasma clots were prepared as described above for turbidity assessment, except that Alexa 488–labeled fibrinogen (Invitrogen, Carlsbad, CA, USA) was added to the plasma before clot formation. Three-dimensional image data sets of clots were collected with a Zeiss LSM510 confocal microscope (Zeiss, Oberkochen, Germany). Reconstructions of 60 z-sections were computed.

Leong L., Chernysh I.N., Xu Y., Sim D., Nagaswami C., de Lange Z., Kosolapova S., Cuker A., Kauser K, & Weisel J.W. (2017). Clot stability as a determinant of effective factor VIII replacement in hemophilia A. Research and Practice in Thrombosis and Haemostasis, 1(2), 231-241.

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Coagulation Assays for Heparin Monitoring

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aPTT was determined using HemosIL aPTT-SP (silicate activator; Instrumentation Laboratory, Bedford, Massachusetts, USA). Fifty microlitres of patient plasma were mixed with 50 µl of aPTT-SP. TT is a coagulation test measuring the time to fibrin formation after adding thrombin.23 Because of a high sensitivity for heparin, two thrombin concentrations are necessary to cover the full spectrum of heparin concentration. We used 3 U/mL (TT 1) and 7.5 U/mL (TT 2) of bovine thrombin (HemosIL TT; Instrumentation Laboratory, Bedford, Massachusetts, USA). TT 2 was determined in case of an unclottable TT 1 corresponding to a high concentration of heparin. Anti-Xa activity was determined using HemosIL liquid anti-Xa (Instrumentation Laboratory, Bedford, Massachusetts, USA), calibrated with HemosIL heparin calibrators. PiCT was measured using Pefakit PiCT (DSM Pentapharm, Basel, Switzerland). The test has been described in detail elsewhere.19 24 (link) Briefly, 50 µL of patient plasma was incubated for 180 s with an activator containing activated factor Xa, phospholipids and Russell’s viper venom. Coagulation was initiated with calcium chloride and clotting time (s) measured. All analyses were performed on an ACL TOP 300 (Instrumentation Laboratory, Bedford, Massachusetts, USA).

Bürki S., Brand B., Escher R., Wuillemin W.A, & Nagler M. (2018). Accuracy, reproducibility and costs of different laboratory assays for the monitoring of unfractionated heparin in clinical practice: a prospective evaluation study and survey among Swiss institutions. BMJ Open, 8(6), e022943.

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Dabigatran Concentration Measurement Protocol

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DC at the trough and peak times was calculated using HemosIL^® direct thrombin inhibitor assay (Instrumentation Laboratory, Bedford, MA, USA). This assay is a dilute thrombin time test, which is based on the reaction between dabigatran and exogenous thrombin added to the diluted patient plasma. The associated clotting time was measured using the ACL TOP hemostasis testing system (Instrumentation Laboratory), and then the concentration of dabigatran was estimated from the reference curve of the known plasma standard of dabigatran using HemosIL^® dabigatran calibrators　(Instrumentation Laboratory). The trough or peak ΔDC ratio was defined as the trough or peak DC in the period without PPI minus the corresponding DC in the period with PPI divided by the trough or peak DC in the period with PPI. Trough and peak aPTTs were measured using HemosIL^® APTT-SP (Instrumentation Laboratory).

Kuwayama T., Osanai H., Ajioka M., Tokuda K., Ohashi H., Tobe A., Yoshida T., Masutomi T., Kambara T., Inoue Y., Nakashima Y., Asano H, & Sakai K. (2017). Influence of proton pump inhibitors on blood dabigatran concentrations in Japanese patients with non-valvular atrial fibrillation. Journal of Arrhythmia, 33(6), 619-623.

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Clot Structure Analysis in Hemophilia A

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Kinetics of turbidity development during activated partial thromboplastin time (aPTT) reactions allowed indirect assessment of clot structure. Plasma samples from individual HemA donors (HRF, Inc, Raleigh, NC, USA) were spiked with FVIII (0‐100%), and the aPTT and fibrinogen assays were performed with the HemosIL APTT‐SP and Fibrinogen‐C kits, respectively (Instrumentation Laboratory [IL], Bedford, MA, USA), using on‐board ACL TOP tests. HemA and control clots were prepared using similar conditions as the aPTT assay using HemA and pooled plasma from healthy donors (HRF, Inc), respectively. CT was determined from the second derivative of the turbidity change over time, and turbidity waveform data were extracted from the ACL TOP using proprietary software developed by Bayer Business Services (Leverkusen, Germany). Changes in turbidity parameters were correlated with changes in CTs and with fibrinogen levels.
Direct visualization of plasma clots was achieved by confocal microscopy of plasma. HemA plasma clots were prepared as described above for turbidity assessment, except that Alexa 488‐labeled fibrinogen (Invitrogen, Carlsbad, CA, USA) was added to the plasma before clot formation. Three‐dimensional image data sets of clots were collected with a Zeiss LSM510 confocal microscope (Zeiss, Oberkochen, Germany). Reconstructions of 60 z‐sections were computed.

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Coagulation Factors Measurement Protocol

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FVIII:C and FIX:C levels were measured using one‐stage clotting assay (HemosIL™ APTT‐SP; Instrumentation Laboratory and Coagpia^® APTT‐N; Sekisui Medical Co.) on an ACL 9000 Automated Coagulometer and CP3000™. VWF:Ag levels were measured using latex coagulating nephelometry (STA Listest vWF[FR]; Diagnostica Stago, Inc.) on a JCA‐BM8020 (JEOL).

Shinozawa K., Amano K., Hagiwara T., Bingo M., Chikasawa Y., Inaba H., Kinai E, & Fukutake K. (2021). Genetic analysis of carrier status in female members of Japanese hemophilia families. Journal of Thrombosis and Haemostasis, 19(6), 1493-1505.

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Chromogenic Anti-Xa Assay for Liquid Heparin Monitoring

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The HemosIL Liquid Heparin Kit (Instrumentation Laboratory, Lexington, KY, USA) was used for the chromogenic AXA assay. This 1-stage assay is based on the reaction between FXa and a synthetic chromogenic substrate. For Liquid Heparin, 10 μl of plasma were mixed with 100 μl chromogenic substrate Baseline, first peak and trough and steady-state trough and peak anti-factor Xa activity (AXA) in 25 patients. The respective measurements were 0.01±0.02 IU/ml, 0.83±0.43 IU/ml, 0.34±0.17 IU/ml, 0.92±0.52 IU/ml, and 1.61±0.62 IU/ml. First trough AXA was significantly lower than steady-state trough AXA, although it was significantly higher than baseline. Similarly, first peak AXA was significantly lower than steady-state peak AXA. OSANAI H et al.
PT was measured using HemosIL RecombiPlasTin (Instrumentation Laboratory), and aPTT was measured using HemosIL APTT-SP (Instrumentation Laboratory).

Osanai H., Ajioka M., Masutomi T., Kuwayama T., Ishihama S., Sakamato Y., Otaka N., Sakaguchi T., Inoue Y., Kanbara T., Nakashima Y., Asano H, & Sakai K. (2015). Measurement of Anti-Factor Xa Activity in Patients on Apixaban for Non-Valvular Atrial Fibrillation. Circulation journal : official journal of the Japanese Circulation Society, 79(12).

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Comparative Evaluation of aPTT Reagents

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We evaluated a single batch of five commercially available aPTT reagents; HemosIL APTT SP (Instrumentation Laboratory, IL, Bedford, MA, USA), HemosIL SynthASil (IL), STA-CK Prest 5 (Stago, Asni eres, France), Trini-CLOT aPTT HS (TCoag, Wicklow, Ireland), and Trini-CLOT Automated aPTT (TCoag). The composition in phospholipids and contact system activator of these reagents was reported in Table 1. The mean control clotting times and normal ranges, as indicated by the manufacturers, are shown in the Table 2.

Toulon P., Eloit Y., Smahi M., Sigaud C., Jambou D., Fischer F, & Appert-Flory A. (2016). In vitro sensitivity of different activated partial thromboplastin time reagents to mild clotting factor deficiencies. International journal of laboratory hematology, 38(4).

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Coagulation Assays in Citrated Plasma

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Determination of APTT and PT times and fibrinogen concentration were measured using commercially available reagents (APTT: HEMOSIL® APTT-SP, Instrumentation Laboratory, Werfen Company, Bradford, USA; PT: RecombiPlasTin 2G, Instrumentation Laboratory, Werfen Company, Bradford, USA; Fibrinogen: Q.F.A. Thrombin (Bovine), Instrumentation Laboratory, Werfen Company, Bradford, USA) in the citrated plasma. In order to determine the APTT, PT and fibrinogen concentration, ACL ELITE PRO was used.

Budkowska M., Lebiecka A., Marcinowska Z., Woźniak J., Jastrzębska M, & Dołęgowska B. (2019). The circadian rhythm of selected parameters of the hemostasis system in healthy people. Thrombosis research, 182.

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