凝血致活酶

摘要： 慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)目前是全世界范围内发病率和死亡率最高的疾病之一,是严重的公共卫生问题.肺动脉高压是COPD常见的并发症.目前关于肺动脉高压的发病机制尚不明确,缺氧在COPD合并肺动脉高压的发病中扮演着重要的角色,而高原地区环境的低氧会加重呼吸系统疾病.有研究发现在肺动脉高压的发生、发展过程中,导致凝血功能改变的众多因素中,炎症因子可能占主导性作用,本文主要探讨肿瘤坏死因子α(TNF-α)、组织因子在肺动脉高压中的发病机制中与凝血功能之间的作用,为高原地区肺动脉高压的发生发展提供新的理论依据.

摘要： 免疫性血小板减少症(ITP)为一种获得性自身免疫性出血性疾病,该病涉及血小板破坏增加与血小板生成不足导致的血小板计数减少,伴或者不伴出血的临床表现.ITP患者血小板计数减少可以导致机体凝血功能障碍引起出血,但是出血情况的严重程度并不完全与血小板计数减少程度呈正相关关系.部分患者血小板计数即使极低亦无明显出血现象,这表明存在额外的机制代偿凝血功能,并改善这些患者的出血情况.笔者拟就近年有关ITP患者凝血代偿机制的研究进展进行综述.%Immune thrombocytopenia (ITP) is an acquired autoimmune hemorrhagic disease involving increased platelet destruction and insufficient platelet production,leading to thrombocytopenia,with or without clinical manifestations of bleeding.Although thrombocytopenia may cause coagulation dysfunction and hemorrhagic complications,the bleeding severity is not certainly proportional to the number of platelets.In some cases,there is no significant bleeding symptom even the platelet count is extremely low,which suggests there are additional compensatory mechanisms to control or ameliorate the bleeding complications in ITP patients.The present article aims to review recent investigations on the compensatory mechanisms of coagulation function in the patients with ITP.

摘要： 子痫前期(PE)是妊娠期常见的并发症,也是导致不良妊娠结局的主要原因,严重者可致母儿死亡.但其发病机制尚不清楚,其发生可能与孕妇年龄、慢性高血压、糖尿病、孕前肥胖和多胎妊娠等有关.PE是一个复杂的多系统疾病,主要涉及胎盘滋养层异常入侵、抗血管生成因子异常表达、氧化应激和炎症反应等.研究发现血栓形成相关因子以及炎症相关因子与PE的发生有关,能够促进病情进展,但具体机制不明.理清血栓形成及炎症在PE发病中发挥的作用可以为PE的早期预防和治疗提供相关策略,现就血栓形成及炎症在PE发生、发展中的作用作一综述.

摘要： [目的]分析脑出血患者血清组织因子途径抑制物(TFPI-1)、组织因子(TF)、纤维蛋白原(FIB)、一氧化氮(NO)表达情况及其临床意义.[方法]选取2015年12月至2018年4月本月收治的120例脑出血患者的临床资料(观察组),根据美国国立卫生研究院卒中量表(NIHSS)评分对其病情严重程度分为轻度组、中度组和重度组,根据血肿体积分为小血肿组、中血肿组和大血肿组.选取同时期于本院进行健康体检的100例健康志愿者作为对照组.采用双抗体夹心酶联免疫吸附法检测血清TFPI-1、TF、FIB、NO水平,比较各组血清TFPI-1、TF、FIB、NO表达.[结果]观察组患者TFPI-1、TF、FIB、NO水平均显著高于对照组,差异有统计学意义(P＜0.05).重度组患者血清TFPI-1、TF、FIB、NO水平高于中度组、轻度组,中度组患者血清TFPI-1、TF、FIB、NO水平高于轻度组,差异均有统计学意义(P＜0.05).大血肿组患者血清TFPI-1、TF、FIB、NO水平高于中血肿组、小血肿组,中血肿组患者血清TFPI-1、TF、FIB、NO水平高于小血肿组,差异均具有统计学意义(P＜0.05).[结论]血清TFPI-1、TF、FIB、NO水平随着血肿量增多和病情进展而升高,提示血清TFPI-1、TF、FIB、NO在一定程度上参与了脑出血的病情发生、发展.

摘要： 目的:观察比较冠心病(CHD)患者和健康居民间同型半胱氨酸(Hcy) 、血管假性血友病因子(vWF)和组织因子促凝活性(TF-PCA)的差异,及3项指标与冠心病发病的关系.方法:回顾性分析2013年4月 -2015年12月期间,我院心内科收治的95例CHD急诊患者(CHD组)和同期行健康体检的95例健康居民(健康对照组)的临床资料.根据病情,CHD组被进一步分为稳定型心绞痛(SAP)组(33例) 、不稳定型心绞痛(UAP)组(31例)和急性心肌梗死(AMI)组(31例).测量比较各组以及CHD患者不同发病时间的 Hcy 、 vWF和TF-PCA,多因素Logistic回归分析冠心病类型的影响因素.结果:与健康对照组比较,CHD组 Hcy [ (9.22 ± 3.45) μmol/L比(17.80 ± 6.94) μmol/L]、 vWF [ (122.40 ± 10.18)% 比(160.13 ± 10.48)%]和TF-PCA [ (30.12 ± 10.49) s比(69.45 ± 8.26) s]均显著升高,P均＝0.001.与SAP组比较,UAP组和AMI组的Hcy [ (14.30 ± 3.15) μmol/L比(20.50 ± 4.97) μmol/L比(25.77 ± 6.10) μmol/L]、 vWF [ (141.56 ± 9.45)% 比(168.23 ± 11.29)% 比(185.56 ± 11.40)%]和TF-PCA [ (45.13 ± 11.52) s比(53.16 ± 18.45) s比(64.49 ± 11.59) s]均显著升高,且AMI组的显著高于UAP组的,P＜0.05或＜0.01.随着发病时间的推移,CHD患者Hcy 、 vWF和TF-PCA水平均显著降低,且均符合1h＞12h＞24h＞48h,两两比较均有显著差异,P均＝0.001.多因素Logistic回归分析显示,Hcy、 vWF和TF-PCA是冠心病类型的独立危险因素(OR＝2.586～5.058,P均＝0.001).结论:冠心病患者Hcy 、 vWF和TF-PCA水平均显著升高,病情越严重,水平越高;随着时间推移,其水平均显著下降.三项指标均为冠心病类型的独立危险因素.%Objective:To observe and compare differences of levels of homocysteine (Hcy),von Willebrand factor (vWF) and tissue factor procoagulant activity (TF-PCA) between patients with coronary heart disease (CHD) and healthy resi-dents,and analyze relationship between above 3 indicators and coronary heart disease onset.Methods:Clinical data of 95 CHD emergency patients (CHD group),who were treated in our department of cardiology from Apr 2013 to Dec 2015,and 95 healthy residents (healthy control group) were retrospectively analyzed.According to state of an illness,CHD group was further divided into stable angina pectoris (SAP) group (n＝33),unstable angina pectoris (UAP) group (n＝31) and acute myocardial infarction (AMI) group (n＝31).Levels of Hcy,vWF and TF-PCA were measured and compared among all groups and different time after onset in CHD patients.Multifactor Logistic regression analysis was used to analyze influ-encing factors of CHD types.Results:Compared with healthy control group,there were significant rise in levels of Hcy [ (9.22 ± 3.45) μmol/L vs.(17.80 ± 6.94) μmol/L],vWF [(122.40 ± 10.18)% vs.(160.13 ± 10.48)%] and TF-PCA [ (30.12 ± 10.49) s vs.(69.45 ± 8.26) s] in CHD group,P＝0.001 all.Compared with SAP group,there were signifi-cant rise in levels of Hcy [ (14.30 ± 3.15) μmol/L vs.(20.50 ± 4.97) μmol/L vs.(25.77 ± 6.10) μmol/L],vWF [ (141.56 ± 9.45)% vs.(168.23 ± 11.29)% vs.(185.56 ± 11.40)%] and TF-PCA [ (45.13 ± 11.52) s vs.(53.16 ± 18.45) s vs.(64.49 ± 11.59) s] in UAP group and AMI group,and those of AMI group were significantly higher than those of UAP group,P＜0.05 or ＜0.01.As onset time went by,there were significant reductions in levels of Hcy,vWF and TF-PCA in CHD patients,and all of them accorded with 1h＞12h＞24h＞48h,there existed significant difference be- tween any two time points,P＝0.001 all.Multifactor Logistic regression analysis indicated that Hcy,vWF and TF-PCA were independent risk factors for CHD type (OR＝2.586～5.058,P＝0.001 all).Conclusion:Levels of Hcy,vWF and TF-PCA significantly rise in CHD patients,the more severe disease is,the higher levels are.Along with time goes by,their levels significantly reduce.Combined detection of them can be used for predicting CHD type.The Hcy,vWF and TF-PCA are independent risk factors for CHD type.

摘要： 目的 观察五步蛇毒蛋白C激活物(PCA)对人脐静脉血管内皮细胞(HUVEC)的保护作用,并分析其对组织因子(TF)表达的影响.方法 常规培养HUVEC,实验分为空白对照组、脂多糖(LPS)组、PCA(0.625、1.25、2.50 μg ·mL-1)组和PCA+LPS组.MTT法检测HUVEC活性,ELISA检测HUVEC培养上清中TF分泌量,免疫荧光染色检测核转录因子(NF)-κB是否被激活-核转运.结果PCA对HUVEC活性和形态无显著影响(P＞0.05).与空白对照组,LPS组细胞活性显著下降(P＜0.01);而LPS+PCA组细胞活性显著高于LPS组(P＜0.05).LPS组TF表达量和NF-κB表达均显著高于空白对照组(P＜0.01);LPS +PCA组与LPS组比较TF表达量和NF-κB表达降低(P＜0.01).结论 PCA可降低HUVEC分泌TF,减少LPS对HUVEC的损伤作用,其可能与抑制NF-κB通路的活化有关.%AIM To observe the protective effects of Agkistrodon acutus venom protein C activator (PCA) on human umbilical vein endothelial cells (HUVEC),and analyze PCA's effects for tissue factors (TF) expression.METHODS HUVEC were conventionally cultured and divided into blank control group,lipopolysaccharide (LPS) group,PCA (0.625,1.25,2.50 μg·mL-1) group and PCA + LPS group.MTT assay was used to detect the HUVEC viability.The secretion level of TF in HUVEC medium was performed by ELISA.Cellular localization of NF-κB was detected with immunofluorescence staining.RESULTS PCA had no significant effect on HUVEC activity and morphology (P ＞ 0.05).The cell viability of LPS group was significantly decreased compared with the blank control group (P ＜ 0.01),and the viability of LPS + PCA group was significantly increased in the LPS + PCA group than in the LPS group (P ＜ 0.05).The expression of TF and NF-κB were significantly increased in the LPS group than in the blank control group (P ＜ 0.01).But the expression of TF and NF-κB was significantly decreased in the LPS + PCA group than in the LPS group (P ＜ 0.01).CONCLUSION PCA can protect HUVEC viability by reducing the secretion of TF in LPS-treated HUVEC,which may be associated with the suppression of NF-κB activation.

摘要： Objective To observe the effect of simvastatin on plasma tissue factor and tissue factor pathway inhibitor in hemodialysis patients,and to explore the value of simvastatin in the treatment of hemodialysis patients.Methods Ninety cases of chronic renal failure patients with hemodialysis treatment were randomly divided into conventional treatment group and simvastatin treatment group, and 50 cases healthy people received in the same period as control group.Plasma tissue factor(TF), tissue factor pathway inhibitor(TFPI) activity and blood lipid level triglyceride(TG), total cholesterol(TC), high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol(LDL-C) changes before and after treatment were compared.Results TF and TFPI activity in chronic renal failure patients were significantly higher than those in control group(P0.05).Pearson correlation analysis showed that TF and TPPI levels were not correlated with TG, TC, HDL-C and LDL-C(P>0.05).Conclusion 20 mg simvastatin treatment in maintenance hemodialysis patients can reduce the tissue factor activity, increase tissue factor pathway inhibitor level, which was important for correcting coagulation disorders in hemodialysis patients.%目的 观察辛伐他汀对血液透析患者血浆组织因子血浆组织因子(tissue factor,TF)和组织因子途径抑制物(tissue factor pathway inhibitor, TFPI)的影响,探讨辛伐他汀在血液透析患者治疗中的应用价值.方法 选择行血液透析治疗的慢性肾功能不全患者90例,依据随机数字法分为常规治疗组和辛伐他汀组,并选取同期体检的50例健康人群作为对照组,比较治疗前后TF、TFPI水平及血脂水平[三酰甘油(triglyceride,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)和低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)]变化.结果 肾功能不全患者TF和TFPI水平明显高于对照组(P0.05).Pearson相关性分析表明,TF、TPPI水平与TG、TC、HDL-C、LDL-C均无相关性(P>0.05).结论 20 mg辛伐他汀治疗维持性血液透析患者,能够降低TF、升高TFPI,对于纠正血液透析患者凝血功能紊乱具有重要意义.

摘要： Objective:To preliminarily verify the cross talk between tissue factor/active coagulation factor Ⅶ (TF/FⅦa) and epidermal growth factor receptor (EGFR) pathways in human colon cancer cells in culture.Methods:FⅦa was treated to HT-29 (KRAS-wild type) and LoVo (KRAS-mutant) colon cancer cells to activate TF/F Ⅶa pathway,qRT-PCR and Western blot were used to detect the expressions of amphiregulin (AREG) and epiregulin (EREG),ligands of EGFR on mRNA and protein levels,respectively.After knocking down expression of TF by TF-targeted siRNA transfection,FⅦa was treated and mRNA expressions of AREG and EREG were detected to see whether the FⅦa-induced effects were dependent on TF.Expressions of mRNA of TF and FⅦwere detected by qRT-PCR following the activation of EGFR pathway by treatment with epidermal growth factor (EGF) to HT-29 and LoVo cells.Results:After TF/FⅦa pathway was activated,for HT-29 cells,expressions of AREG (on mRNA level) and EREG (both on mRNA and protein level) were significantly down-regulated versus those of control group,gene expressions of AREG and EREG were 0.55 ± 0.09 vs.0.99 ± 0.09,0.67 ± 0.10 vs.1.02 ± 0.02,protein expressions of EREG were 0.54 ± 0.09 vs.1.04 ± 0.13,all P ＜ 0.05.For LoVo cells,expressions of AREG (both on mRNA and protein level) and EREG (on protein level) were significantly up-regulated versus those of control group,gene expression of AREG were 1.87 ± 0.39 vs.0.93 ± 0.23,protein expressions of AREG and EREG were 3.09 ±0.73 vs.1.11 ±0.21,1.53 ±0.19 vs.0.97 ± 0.23,all P ＜0.05.The regulating effect of AREG and EREG mRNA expression by FⅦa in HT-29 and LoVo cells could both be partly blocked by knocking down TF expression.For HT-29 cells,activation of EGFR pathway induced no significant TF mRNA expression,F Ⅶ mRNA expression was not detected.However,for LoVo cells,activation of EGFR pathway induced significantly higher mRNA expressions of both TF and FⅦ,expressions were 1.53 ± 0.23 vs.1.00 ± 0.23,53.20 ± 6.08 vs.1.00 ± 0.15,all P ＜0.05.Conclusion:In colon cancer cell LoVo,when activated,TF/FⅦa pathway and EGFR pathway could interact through upregulating the other pathway's effectors,and mutant KRAS might play a critical role in the two pathways'cross talk.%目的:探讨结肠癌细胞中组织因子/活性凝血因子Ⅶ(tissue factor/active coagulation factorⅦ,TF/FⅦa)通路与表皮生长因子受体(epidermal growth factor receptor,EGFR)通路之间是否存在交互作用.方法:在KRAS野生型的HT-29及KRAS突变型的LoVo结肠癌细胞中,以FⅦa活化TF/FⅦa通路,采用qRT-PCR、Western blot检测EGFR配体双调蛋白(amphiregulin,AREG)及表皮调节素(epiregulin,EREG)基因、蛋白表达改变;利用RNA干扰技术敲低TF表达后活化TF/FⅦa通路,检测其对AREG、EREG基因表达的影响,以证实FⅦa对AREG、EREG表达调节作用依赖TF.以表皮生长因子(epidermal growth factor,EGF)激活两细胞EGFR通路后,检测TF/FⅦa通路关键分子TF、FⅦ基因表达改变.结果:TF/FⅦa通路活化后,HT-29细胞AREG、EREG基因表达及EREG蛋白表达水平均较对照组显著下调(AREG、EREG基因表达量分别为0.55±0.09 vs.0.99±0.09、0.67±0.10vs.1.02 ±0.02,EREG蛋白表达量0.54±0.09 vs.1.04±0.13,p均＜0.05);LoVo细胞AREG基因表达及AREG、EREG蛋白表达水平较对照组显著上调(AREG基因表达量1.87±0.39 vs.0.93±0.23,AREG、EREG蛋白表达量3.09 ±0.73 vs.1.11 ±0.21、1.53±0.19 vs.0.97±0.23,P均＜0.05);TF敲低后均可部分阻断FⅦa对两细胞AREG、EREG表达调节作用;EGFR通路激活后,HT-29细胞TF基因表达较对照组无显著变化,FⅦ基因表达未检测到,而LoVo细胞的FⅦ及TF基因表达均较对照组显著上调,表达量分别为1.53±0.23 vs.1.00±0.23、53.20±6.08 vs.1.00±0.15(P均＜0.05).结论:结肠癌LoVo细胞TF/FⅦa通路与EGFR通路的活化可分别上调另一通路的关键分子表达并发生交互作用,KRAS基因突变可能对该交互作用的发生发挥关键作用.

摘要： 目的探讨低分子肝素（LMwH）联合5-氟尿嘧啶（5-FU）对荷H22肝癌小鼠血液中组织因子（TF）及组织因子途径抑制物（TFPI）浓度的影响。方法选择40只昆明小鼠，随机分为空白对照组、荷瘤组、LMwH组、LMWH联合5-Fu组（联合组）和5-FU组，各8例。制备H。肝癌荷瘤小鼠模型（空白对照组除外），各组分别予以生理盐水、生理盐水、LMWH、LMWH＋5-FU、5-Fu治疗2周。治疗结束后，经心脏取血，应用ELISA法检测各组血液中TF、TFPI的浓度，并进行比较。结果荷瘤组小鼠血液TF浓度高于空白对照组，TFPI浓度低于空白对照组；LMWH组、联合组及5-FU组小鼠血液中TF的浓度均低于荷瘤组，TFPI的浓度高于荷瘤组，差异有显著性（F=82．534、34．755，P〈0．05）。LMWH组、5-Fu组、联合组小鼠血液中TF、TFPI浓度比较，差异无统计学意义（P〉0．05）。结论LMWH与5-Fu均对肝癌小鼠TF有抑制作用，两者都可提高TFPI含量，但二者无协同作用。

摘要： 本发明涉及一种用于抗凝血酶III活性测定的凝血酶发色底物液、一种用于抗凝血酶III活性测定的凝血酶水溶液、一种用于确定样本中抗凝血酶III活性的方法、一种用于抗凝血酶III活性测定的试剂盒、山梨醇在制备凝血酶发色底物液中的用途以及蔗糖在发色底物法测定抗凝血酶III活性中的用途。用于抗凝血酶III活性测定的凝血酶发色底物液，含有：凝血酶发色底物；山梨醇；以及水。该凝血酶发色底物液能有效地用于测定抗凝血酶III活性。

摘要： 本发明涉及一种活化部分凝血活酶时间测定试剂卡，所述试剂卡包括第一反应区和第二反应区，所述第一反应区和第二反应区分别为将第一反应试剂和第二反应试剂分别均匀涂布于基材表面，干燥后得到；通过采用鞣花酸和白陶土这两种激活剂分别与适宜浓度的羟丙基甲基纤维素、无机盐配制形成两种反应试剂，并涂布在基材表面干燥后制备而成一种用于检测APTT的干式试剂卡，其对肝素敏感性高，检测结果更准确，从而可有效应用于临床肝素治疗评价中，并避免了市售常规冻干粉或液体试剂，由于剂型原因，对保存条件要求高的情况，使用更方便，稳定性强，更利于试剂在实际临床检测中的广泛应用，为实际APTT的检测提供便利。

摘要： 本发明公开了一种用于测定活化部分凝血活酶时间的体外诊断试剂盒，包括APTT检测试剂和用于盛放所述APTT检测试剂的塑料瓶；所述APTT检测试剂由部分凝血活酶试剂和钙盐溶液组成，所述部分凝血活酶试剂由脑磷脂溶液和缓冲液配制而成，所述缓冲液由下述原料制成：活化剂，tris，氯化钠，甘露醇，聚乙二醇，牛血清白蛋白，丙氨酸，石炭酸，proclin300和水。本发明的用于测定活化部分凝血活酶时间的体外诊断试剂盒，使用方便，在保证APTT检测试剂的准确性和灵敏性的基础上，提高APTT检测试剂的稳定性，使该体外诊断试剂盒在临床上广泛应用，使APTT检验实现标准化。

摘要： 本申请公开了一种提高活化部分凝血活酶时间试剂对肝素敏感性的方法及用途。本申请的方法，包括在采用活化部分凝血活酶时间试剂对血浆进行检测时，向反应体系中加入可溶性锰离子和/或镁离子金属盐；或者预先在活化部分凝血活酶时间试剂中添加可溶性锰离子和/或镁离子金属盐，再用于活化部分凝血活酶时间检测。本申请的方法，通过添加可溶性锰离子和/或镁离子的金属盐，进而提高了活化部分凝血活酶时间试剂对肝素的敏感性，使得活化部分凝血活酶时间试剂能够更灵敏的反映肝素浓度变化。本申请的用途包括可溶性锰离子或镁离子金属盐在制备对肝素敏感的活化部分凝血活酶时间试剂中的用途。

摘要： 本发明提供活化部分凝血活酶时间测定方法。此方法包括：将被检血浆和含活化剂及25μg/mL以上的磷脂酰甘油的第1试剂混合的第1混合步骤,将第1混合步骤中得到的样品和含钙盐的第2试剂混合的第2混合步骤，和测定第2混合步骤中得到的样品的凝固时间的步骤。

摘要： 一种凝血酶溶液、试剂盒、凝血酶的稳定化方法、检测试剂以及凝血酶时间的测定方法。凝血酶溶液含有凝血酶，并含有消泡剂。该凝血酶溶液具有提高的稳定性，使得凝血酶在活性较低的情况下也能够保持较长时间的稳定。

摘要： 本发明涉及一种用于抗凝血酶III活性测定的凝血酶发色底物液、一种用于抗凝血酶III活性测定的凝血酶水溶液、一种用于确定样本中抗凝血酶III活性的方法、一种用于抗凝血酶III活性测定的试剂盒、山梨醇在制备凝血酶发色底物液中的用途以及蔗糖在发色底物法测定抗凝血酶III活性中的用途。用于抗凝血酶III活性测定的凝血酶发色底物液，含有：凝血酶发色底物；山梨醇；以及水。该凝血酶发色底物液能有效地用于测定抗凝血酶III活性。

摘要： 本发明涉及获得单体形式的重组凝血酶原激活蛋白酶(rLopap)的方法；重组凝血酶原激活蛋白酶(rLOPAP)及其氨基酸序列。此外，本发明还涉及这种蛋白酶用于消除血液纤维蛋白原以及用作凝血酶原血异常诊断试剂盒的用途。本发明描述了21kDa的重组形式的凝血酶原激活蛋白酶的获得和表征，该蛋白酶被命名为rLopap(Lonomiaobliqua凝血酶原激活蛋白酶)，具有丝氨酸蛋白酶特征，不过它显示出如同lipocalin家族的保守氨基酸序列。该蛋白表现出促凝血活性，消除血液中的血纤维蛋白原并延长人血液/血浆的凝固时间。本发明介绍了重组形式的rLopap的获得，该rLopap显示出足够的活性以便进行临床药理学测定。

摘要： 一种检测凝血酶、类凝血酶及凝血酶抑制剂的纤维蛋白原平板方法，使用琼脂糖与纤维蛋白原制作成纤维蛋白原琼脂平板，通过标准品及检品在该平板上的扩散反应作定量检测。本发明的检测凝血酶、类凝血酶及凝血酶抑制剂的纤维蛋白原平板法，操作简便、直观、稳定、敏感度高(检测到每毫升0.15酶活力单位)，不受检品浊度或颜色干扰，成本低廉。在普通实验室中只要有恒温箱即可实施，是一种易于推广使用的实验方法。

摘要： 本发明涉及一种在临床上同时对五种出血性疾病进行检测的方法和试剂盒的制作。其特征包括以下步骤:步骤（1）制作含有新鲜混合血浆、硫酸钡吸附血浆和储存血清的试剂盒。步骤（2）在试剂盒的每个管中加入一定量待测患者血浆，分别对上述各管测定活化部分凝血活酶时间（APTT）。通过含试剂管与患者血浆APTT时间的对比情况(纠正情况)对五种出血性疾病进行实验室诊断。步骤（3）用储存血清、硫酸钡吸附血浆分别对混合血浆进行不同浓度稀释后，测定活化部分凝血活酶时间（APTT）得到因子Ⅷ、Ⅸ标准曲线，对患者因子Ⅷ、Ⅸ进行定量测定。