High Sensitive ELISA Kit for Prothrombin Fragment 1+2 (F1+2)

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Prothrombin Fragment 1+2 (F1+2).
No significant cross-reactivity or interference between High Sensitive Prothrombin Fragment 1+2 (F1+2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Prothrombin Fragment 1+2 (F1+2) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Prothrombin Fragment 1+2 (F1+2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 81-99 95
EDTA plasma(n=5) 90-101 93
heparin plasma(n=5) 89-96 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Prothrombin Fragment 1+2 (F1+2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Prothrombin Fragment 1+2 (F1+2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive Prothrombin Fragment 1+2 (F1+2) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-103% 91-105% 83-90% 93-105%
EDTA plasma(n=5) 91-101% 85-99% 82-89% 87-94%
heparin plasma(n=5) 97-105% 79-98% 90-104% 78-101%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Critical Care Disseminated intravascular coagulation or acute coagulopathy of trauma shock early after trauma? An observational study BioMed: cc10553
Journal of Vascular Surgery Changes in thrombin generation, fibrinolysis, platelet and endothelial cell activity, and inflammation following endovascular abdominal aortic aneurysm repair ScienceDirect: S0741521411018568
Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine High levels of soluble VEGF receptor 1 early after trauma are associated with shock, sympathoadrenal activation, glycocalyx degradation and inflammation in severely injured patients: a prospective study Sjtrem:1757-7241
Journal of Thrombosis and Haemostasis High sCD40L levels early after trauma are associated with enhanced shock, sympathoadrenal activation, tissue and endothelial damage, coagulopathy and mortality Wiley: source
British Journal of Dermatology Evaluation of autologous plasma skin test in patients with chronic idiopathic urticaria PubMed: 21910697
Arteriosclerosis, Thrombosis, and Vascular Biology. Diet Modulates Endogenous Thrombin Generation, A Biological Estimate of Thrombosis Risk, Independently of the Metabolic Status PubMed: 22859493
Journal of Vascular Surgery Medium-term effect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysis PubMed: 23140799
PLoS ONE Markers of Thrombogenesis and Fibrinolysis and Their Relation to Inflammation and Endothelial Activation in Patients with Idiopathic Pulmonary Arterial Hypertension Plosone: Source
Journal of Vascular Surgery Effect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysis Pubmed: 23140799
Haemetology Clinical impact of factor V Leiden, prothrombin G20210A, and MTHFR C677T mutations among sickle cell disease patients of Central India Pubmed: 23992124
Cogent Medicine Study the association of adiponectin with inflammation and hypercoagulability in case of type 2 diabetic subjects with renal dysfunction Tandfonline:Source
Interact Cardiovasc Thorac Surg. An ex vivo evaluation of blood coagulation and thromboresistance of two extracorporeal circuit coatings with reduced and full heparin dose Pubmed:24632424
Perfusion Ex vivo simulation of cardiopulmonary bypass with human blood for hemocompatibility testing PubMed: 26243277
J Thromb Haemost Comparison of the Effect of Dabigatran and Dalteparin on Thrombus Stability in a Murine Model of Venous Thromboembolism PubMed: 26514101
Acta Oncologica Turcica Thrombin activatable fibrinolysis inhibitor (TAFI), tissue factor pathway inhibitor (TFPI), and prothrombin fragment 1+2 levels in patients with advanced colorectal cancer aot:AOT_49_1_6_12.pdf
Advances in Medical Sciences Perioperative thrombocytopenia predicts poor outcome in patients undergoing transcatheter aortic valve implantation pubmed:29145170
Thrombosis Research The causes of thrombocytopenia after transcatheter aortic valve implantation pubmed:28582640
Acta Haematologica Elevations of Thrombotic Biomarkers in Hemoglobin H Disease Pubmed:29402840
Journal of Neurotrauma The cerebral thrombin system is activated after intracerebral hemorrhage and contributes to secondary lesion growth and poor neurological outcome in C57Bl/6 mice Pubmed: 31830857
Indian Journal of Public Health Research & Development Enhanced Fibrin-Lysis in Grade-1 Dengue Haemorrhagic Fever
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