Coltest

Manufactured by Roche

Sourced in Germany, Hungary

COLtest is a laboratory instrument designed for the automated analysis of cholesterol levels in biological samples. The device utilizes an enzymatic colorimetric method to quantify the concentration of cholesterol in a specimen.

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Lab products found in correlation

Adptest, by Roche (4 mentions) Traptest, by Roche (3 mentions) Aspitest, by Roche (2 mentions) Hirudin blood tube, by Roche (2 mentions) Multiplate platelet function analyzer, by Roche (1 mentions) Half area 96 well microtiter plates, by Greiner (1 mentions) Victor x5, by PerkinElmer (1 mentions) Sodium chloride (nacl), by B. Braun (1 mentions) Kp 13, by Bachem (1 mentions) Streptozotocin (stz), by Avantor (1 mentions)

7 protocols using coltest

In Vitro Platelet Aggregation Assay

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In vitro aggregation tests were conducted using freshly collected whole blood with Multiplate platelet function analyzer (Roche Diagnostics Polska Sp. z o.o., Warsaw, Poland), the five-channel aggregometer based on measurements of electric impedance. The Multiplate analyzer allows the duplicate measurement with dual electrode probes. Blood was drawn from carotid of rats with hirudin blood tube (Roche Diagnostic). 300 μL of hirudin anticoagulated blood was mixed with 300 μL pre-warmed isotonic saline solution containing studied compound in DMSO or DMSO (0.1% final) and pre-incubated for 3 min at 37 °C with continuous stirring. The agonists (ADPtest, COLtest, Roche Diagnostic) were diluted using isotonic sterile NaCl solution. Aggregation was induced by adding collagen (final concentration 1.6 µg/mL), or adrenaline and subthreshod concentration of ADP (final concentration 50 µM + 1.6 µM). Activated platelet function was recorded for 6 min. The Multiplate software analyzed the area under the curve of the clotting process of each measurement and calculated the mean values.
Data were presented as Mean ± SEM. Statistical comparisons were made by the analysis of variance (ANOVA) and significance of the differences between control group and treated groups was determined by Dunnet post hoc test. p < .05 was considered significant.

Marcinkowska M., Kotańska M., Zagórska A., Śniecikowska J., Kubacka M., Siwek A., Bucki A., Pawłowski M., Bednarski M., Sapa J., Starek M., Dąbrowska M, & Kołaczkowski M. (2018). Synthesis and biological evaluation of N-arylpiperazine derivatives of 4,4-dimethylisoquinoline-1,3(2H,4H)-dione as potential antiplatelet agents. Journal of Enzyme Inhibition and Medicinal Chemistry, 33(1), 536-545.

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Whole Blood Platelet Aggregation Analysis

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The whole blood aggregation was measured using an impedance aggregometer (Multiplate analyzer, Dynabyte Medical, Germany). During the analysis, the sample-reagent mixture was stirred with a discardable PTFE- (polytetrafluoroethylene-) coated magnetic stirrer (800 U/min). Preheated to 37°C saline (300 μl) was placed into the test cells with the addition of anticoagulated whole blood (300 μl) and stirred at 37°C; the measurements were started by adding 20 μl of the appropriate agonist: thrombin receptor activating peptide (TRAP-test, Cat. number 6675883190, Roche Diagnostics) at a final concentration of 10.5 μM, ADP (ADP-test, Cat. number 6675794190, Roche Diagnostics) at a final concentration of 3.2 μM, and collagen (COL-test, Cat. number 6675832190, Roche Diagnostics) at a final concentration of 1.05 μg/ml. The change in impedance by the adhesion and aggregation of platelets on the electrode wires was continuously detected. The mean values of the two determinations are expressed in arbitrary units (AU).

Rola P., Doroszko A., Szahidewicz-Krupska E., Rola P., Dobrowolski P., Skomro R., Szymczyszyn A., Mazur G, & Derkacz A. (2017). Low-Level Laser Irradiation Exerts Antiaggregative Effect on Human Platelets Independently on the Nitric Oxide Metabolism and Release of Platelet Activation Markers. Oxidative Medicine and Cellular Longevity, 2017, 6201797.

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Impedance Aggregometry of Equine Platelets

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Hirudin blood samples were analyzed by the use of impedance aggregometry as described previously.^{40 (link)} Platelet aggregation was measured using 4 different agonists to activate platelets according to the previous manufacturer’s (Dynabyte) information: adenosine diphosphate (6.5 µM final concentration, ADPtest; Roche), ADP + prostaglandin E1 (6.5 µM/9.4 nM final concentration, ADPtestHS; Roche), arachidonic acid (0.75 mM final concentration, ASPItest; Roche), and collagen (1.6 µg/mL final concentration, COLtest; Roche). Spontaneous platelet aggregation (SPA) was also measured by adding isotonic saline instead of a platelet agonist. In 42 control equids and in all SIRS patients, aggregation was measured both with and without stirring for 3 min with a magnetic bar.

Ehrmann C., Engel J., Moritz A, & Roscher K. (2020). Assessment of platelet biology in equine patients with systemic inflammatory response syndrome. Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc, 33(2), 300-307.

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Multiplate Aggregometric POCT Analysis

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Aggregometric POCT in hirudin-anticoagulated whole blood was performed on the multiple electrode aggregometry (MEA) platelet function analyser (Multiplate^®; Roche Diagnostics GmbH, Germany). Test cells of this device incorporate a duplicate sensor for acceptance sampling. Platelets were stimulated in various ways: (1) via arachidonic acid (ASPI test; Roche Diagnostics GmbH, Germany), (2) the ADP receptor (ADP test, Roche Diagnostics GmbH, Germany), (3) the thrombin receptor under the use of thrombin receptor-activating peptide-6 (TRAP test; Roche Diagnostics GmbH, Germany) and (4) via the collagen receptor (COL test, Roche Diagnostics GmbH, Germany). Once initiated, platelet aggregation was allowed to run up to 6 min, and the area under the curve (AUC) was calculated.

Schmitt F.C., Manolov V., Morgenstern J., Fleming T., Heitmeier S., Uhle F., Al-Saeedi M., Hackert T., Bruckner T., Schöchl H., Weigand M.A., Hofer S, & Brenner T. (2019). Acute fibrinolysis shutdown occurs early in septic shock and is associated with increased morbidity and mortality: results of an observational pilot study. Annals of Intensive Care, 9, 19.

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Collagen-Induced Platelet Aggregation Assay

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Platelet-poor plasma (PPP) was prepared from the remaining sample after removal of PRP by centrifugation at 10.000×g for 10 minutes. PRP was pre-incubated with drug or vehicle. Half-area 96-well microtiter plates (Greiner Bio-One, Stonehouse, Gloucestershire,UK) were coated with 5 µL PBS with or without the addition of collagen (COLTest, Roche Diagnostics, Mannheim, Germany) as previously described [29] . The final concentrations of collagen were: 30.0, 12.8, 6.4, 3.2, 1.1, 0.36 and 0.12 µg/mL. Then, 45 µL PRP was added to each well. As reference samples, PRP and PPP were added to wells coated with only PBS. Next, microtiter plates were shaken at 900 rpm for 10 min at 37°C on the Victor X5 (Perkin Elmer, Turku, Finland) with subsequent readings of absorbance and reporting as optical density (OD). Percent of platelet aggregation was calculated using the formula [29] : 100% × (ODPRP -ODsample) / (ODPRP -ODPPP)

Knudsen G.H., Nielsen C., Nielsen C.B., Frederiksen H, & Vinholt P.J. (2020). The effect of mycophenolate mofetil on platelet function. Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis, 31(2).

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Platelet Aggregation by Impedance

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Platelet aggregation was measured by impedance technology (Multiplate, Roche Diagnostics, Basel, Switzerland). The platelet concentration of the sample was adjusted to 300 × 10 9 /L by dilution with plasma from an AB Rh D+ donor (stored at -70ºC until use). To the test cuvette, 300 µL of pre-warmed 9 mg/mL NaCl (B. Braun, Melsungen, Germany) supplemented with 3.0 mM CaCl 2 was added and 300 µL of diluted sample. After 3 min of incubation with constant stirring, 20 µL of platelet agonist (TRAP test, final concentration 32 µM and COLtest, final concentration 3.2 µg/mL, Roche Diagnostics) was added and the aggregation response followed for 6 min at 37ºC under constant stirring.

Tynngård N., Alshamari A., Sandgren P., Kenny D., Vasilache A.M., Abedi M.R, & Ramström S. (2023). High fragmentation in platelet concentrates impacts the activation, procoagulant, and aggregatory capacity of platelets. Platelets, 34(1).

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Platelet Aggregation Assay Protocol

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Hirudin blood tubes, disposable multiplate test cells, and inducers of platelet aggregation (ADPItest, ASPItest [arachidonic acid] and COLtest) were purchased from Roche Ltd. (Budapest, Hungary). KP-13 and RF-9 were purchased from Bachem (Bubendorf, Switzerland). Streptozotocin was obtained from Amresco (Solon, Ohio, USA). The original reagents for the determination of hematological parameters were purchased from Sysmex Ltd. (Budapest, Hungary). High-density lipoprotein (HDL) and triglyceride tests were purchased from DiaSys Diagnostic Systems GmbH (Holzheim, Germany), while the other tests for determining the chemical parameters originate from Roche Ltd. (Budapest, Hungary).

Mezei Z., Váczi S., Török V., Stumpf C., Ónody R., Földesi I, & Szabó G. (2017). Effects of kisspeptin on diabetic rat platelets. Canadian journal of physiology and pharmacology, 95(11).

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