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凝血酶原片段F1+2(F1+2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

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

    • 凝血酶原片段F1+2(F1+2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
    • 凝血酶原片段F1+2(F1+2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
    • 凝血酶原片段F1+2(F1+2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)實(shí)驗(yàn)結(jié)果圖
    • SEA710Hu.jpg標(biāo)準(zhǔn)曲線圖
    • Certificate通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

    特異性

    本試劑盒用于檢測(cè)凝血酶原片段F1+2(F1+2),經(jīng)檢測(cè)與其它相似物質(zhì)無明顯交叉反應(yīng)。
    由于受到技術(shù)及樣本來源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

    回收率

    分別于定值血清及血漿樣本中加入一定量的凝血酶原片段F1+2(F1+2)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

    樣本回收率范圍(%)平均回收率(%)
    serum(n=5)78-9993
    EDTA plasma(n=5)96-105101
    heparin plasma(n=5)97-104101

    精密度

    精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
    批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
    批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
    批內(nèi)差: CV<10%
    批間差: CV<12%

    線性

    在定值血清及血漿樣本內(nèi)加入適量的凝血酶原片段F1+2(F1+2),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中凝血酶原片段F1+2(F1+2)含量的測(cè)定值與理論值的比率。

    樣本1:21:41:81:16
    serum(n=5)90-99%78-103%85-102%98-105%
    EDTA plasma(n=5)88-101%83-104%84-98%84-101%
    heparin plasma(n=5)80-99%84-99%84-101%88-102%

    穩(wěn)定性

    經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
    為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來進(jìn)行操作可減少人為誤差。

    實(shí)驗(yàn)流程

    1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
    2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
    3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
    4. 洗板3次;
    5. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
    6. 洗板5次;
    7. 加TMB底物90µL,37°C孵育10-20分鐘;
    8. 加終止液50µL,立即450nm讀數(shù)。

    實(shí)驗(yàn)原理

    將F1抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標(biāo)準(zhǔn)品或標(biāo)本,其中的F1與連接于固相載體上的抗體結(jié)合,然后加入生物素化的F2抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的F1+2呈正相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

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    參考文獻(xiàn)

    雜志參考文獻(xiàn)
    Critical CareDisseminated intravascular coagulation or acute coagulopathy of trauma shock early after trauma? An observational study[BioMed: cc10553]
    Journal of Vascular SurgeryChanges 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 MedicineHigh 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 HaemostasisHigh 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 SurgeryMedium-term effect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysis[PubMed: 23140799]
    PLoS ONEMarkers of Thrombogenesis and Fibrinolysis and Their Relation to Inflammation and Endothelial Activation in Patients with Idiopathic Pulmonary Arterial Hypertension[Plosone: Source]
    Journal of Vascular SurgeryEffect of endovascular and open abdominal aortic aneurysm repair on thrombin generation and fibrinolysis[Pubmed: 23140799 ]
    HaemetologyClinical impact of factor V Leiden, prothrombin G20210A, and MTHFR C677T mutations among sickle cell disease patients of Central India[Pubmed: 23992124]
    Cogent MedicineStudy 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]
    PerfusionEx vivo simulation of cardiopulmonary bypass with human blood for hemocompatibility testing[PubMed: 26243277]
    J Thromb HaemostComparison of the Effect of Dabigatran and Dalteparin on Thrombus Stability in a Murine Model of Venous Thromboembolism[PubMed: 26514101]
    Acta Oncologica TurcicaThrombin 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 SciencesPerioperative thrombocytopenia predicts poor outcome in patients undergoing transcatheter aortic valve implantation[pubmed:29145170]
    Thrombosis ResearchThe causes of thrombocytopenia after transcatheter aortic valve implantation[pubmed:28582640]
    Acta?HaematologicaElevations of Thrombotic Biomarkers in Hemoglobin H Disease[Pubmed:29402840]
    Journal of NeurotraumaThe 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 & DevelopmentEnhanced Fibrin-Lysis in Grade-1 Dengue Haemorrhagic Fever[]
    Rheumatol IntThe impact of anti-endothelial cell antibodies (AECAs) on the development of blood vessel damage in patients with systemic lupus erythematosus: the preliminary?…[Pubmed:35284968]