Multiplex Assay Kit for Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-104 81
EDTA plasma(n=5) 78-91 84
heparin plasma(n=5) 78-97 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Prothrombin Fragment 1+2 (F1+2) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 78-103% 95-102% 83-99% 90-97%
EDTA plasma(n=5) 94-102% 81-89% 89-97% 95-102%
heparin plasma(n=5) 78-99% 88-95% 96-104% 80-98%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:F1+2) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
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Indian Journal of Public Health Research & Development Enhanced Fibrin-Lysis in Grade-1 Dengue Haemorrhagic Fever
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