CN112941143A - 一种简单便捷的检测凝血和血小板功能的试剂及其应用 - Google Patents
一种简单便捷的检测凝血和血小板功能的试剂及其应用 Download PDFInfo
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Abstract
本发明提供一种检测凝血和血小板功能的试剂及其应用，所述试剂是玻璃珠，所述玻璃珠粒径分布在0.1‑60μm，单个玻璃珠的重量为0.001‑0.01g。该试剂中的玻璃珠是内源性凝血途径中的表面激活物，所述玻璃珠带负电荷当与血液接触时，首先凝血因子FⅩⅡ结合到玻璃珠表面并被激活为FⅩⅡa。FⅩⅡa的主要功能是激活FⅩⅠ成为FⅩⅠa，从而启动内源性凝血途径。所述试剂适用于Sonoclot凝血和血小板功能分析仪。该试剂及其应用以玻璃珠为激活剂，通过体外激活凝血的方法可以对血液样本的凝血及血小板功能进行检测，对早期发现患者是否有血栓形成的危险、提供输血指征，指导血制品的使用、提高手术中及手术后对凝血异常的鉴别与治疗有着重要的意义。
Description
技术领域
本发明涉及医疗器械技术领域，具体地，涉及一种检测凝血和血小板功能的试剂及其应用。
背景
血液凝固是人体内复杂的系统性的生化反应和代谢的过程，其原理是一些的凝血因子协调作用激活凝血酶并将液态的纤维蛋白原转化成固态的纤维蛋白单体的过程。凝血检测，是一种体外检测手段，通过凝血仪和凝血检测试剂对人体的凝血功能进行评价，在临床上主要用于创口手术的风险评估和血栓疾病的诊断以及临床用药的指导。
现场快速检验(point-of-care testing，POCT)是指在采样现场进行的、利用便携式分析仪器及配套试剂快速得到检测结果的一种检测方式。现场快速检验含义可从两方面进行理解：空间上，在患者身边进行的检验，即“床旁检验”，时间上，可进行“即时检验”。近年来，由于当今高新技术的发展和医学科学的进步，以及高效快节奏的工作方式，使得具有实验仪器小型化、操作简单化、报告结果即时化的POCT越来越受到了人们的青睐。但是，现有绝大多数凝血分析是采血后送到实验室由自动化凝血仪进行检测的，周转时间长，送检和存储不当容易影响标本质量。特别是在目前的体外研究中，为了使血液样本起到抗凝的作用，往往要向血液样本中添加抗凝剂肝素。针对添加肝素的这些血液样本，血小板功能的监测是相对困难的，相对缺乏相应的现场快速检测技术支持与临床检测。
现有凝血及血小板功能检测试剂盒，因为试剂配方成份较多，在大规模制作时不可控因素更多，在临床应用中出现的缺点是批次间稳定性差，所以研发一种其效果更优、稳定性更好、配方简单的试剂盒以弥补当前同类试剂盒的不足。
发明内容
发明目的：本发明的目的在于提供一种检测凝血和血小板功能的试剂及其应用，所述试剂可以针对低分子肝素血液样本及自然全血样本的监测，可以广泛应用于人体添加肝素血液样本的枸橼酸钠抗凝全血及自然全血标本的凝血及血小板功能检测中，能够反映动态的凝血全貌，并能具体体现参数ACT(激活全血凝血时间)、CR(凝血速率)及PF血小板功能对围术期的凝血进行监控，提高手术中及手术后对凝血异常的鉴别与治疗。
技术方案：本发明提供一种检测凝血和血小板功能的试剂，所述试剂是玻璃珠，所述玻璃珠粒径分布在0.1-60μm，单个玻璃珠的重量为0.001-0.01g。
进一步的，所述玻璃珠是球形玻璃珠。
进一步的，所述玻璃珠中二氧化硅的含量>90％。
进一步的，所述玻璃珠的D50为15-25μm。
本发明还提供了一种试剂盒，其包括所述的检测凝血和血小板功能的试剂。
所述的检测凝血和血小板功能的试剂、或所述的试剂盒在添加肝素血液样本的枸橼酸钠抗凝全血检测中的应用。
进一步的，所述添加肝素，是每毫升添加0-2单位。
所述的检测凝血和血小板功能的试剂、或所述的试剂盒在自然全血样本检测中的应用。
所述的检测凝血和血小板功能的试剂、或所述的试剂盒在Sonoclot检测仪检测中的应用。
本发明所述试剂中的玻璃珠是内源性凝血途径中的表面激活物，所述玻璃珠带负电荷当与血液接触时，首先凝血因子FⅩⅡ结合到玻璃珠表面并被激活为FⅩⅡa。FⅩⅡa的主要功能是激活FⅩⅠ成为FⅩⅠa，从而启动内源性凝血途径。
本发明所述试剂及其应用，是以玻璃珠为激活剂通过体外激活凝血的方法可以对血液样本的凝血及血小板功能进行检测，该试剂能适用于Sonoclot检测仪，提供激活全血凝血时间(ACT)、凝血速率(CR)及血小板功能(PF)等凝血相关指标，对早期发现患者是否有血栓形成的危险、提供输血指征，指导血制品的使用、提高手术中及手术后对凝血异常的鉴别与治疗有着重要的意义。
有益效果：与现有技术相比，本发明具有以下优势：
1.与进口试剂gbACT+对比结果偏差在5％内，本发明配方各项指标变异系数整体比gbACT+小，稳定性更好，是一种凝血和血小板功能检测的优选试剂配方。
2.所述试剂配方其另一个优点是:试剂配方简单，且成分只有一种就是玻璃珠，容易配置、成本低廉、易规模化生产，是一种替代进口试剂的优选方案。
3.所述方法样本用量小，可针对婴幼儿采血少的问题。
4.所述试剂应用，操作流程简单、人员只要经过简单的培训就可以独立完成操作、检测时间更短。
5.所述试剂配方中的玻璃珠为球形玻璃珠，且此玻璃珠分布均匀，表面光滑，杂质少，所以试剂的稳定性更好。
附图说明
图1为试剂中玻璃珠的粒径分布范围。
图2为本试剂中玻璃珠的形状。
图3为同类型试剂中玻璃珠的形状。
图4为检测结果曲线，反映凝血全过程的图像。
具体实施方式
以下对本发明的配方和方式全面的描述，所数的实施案例是本发明中最优选实施方式，但本发明并不限于以下实施例。
实施例
试剂制备：
(Ⅰ)用定制的称量勺精确称取0.01g球形玻璃珠，此称量勺的过载量仅为1％，能够快速的称量试剂从而提高称量效率。
(Ⅱ)所述球形玻璃珠，中位粒径为15.21μm，单个重量为0.01g，粒径分布在0.1-60μm，其中二氧化硅的含量>90％。
样本准备：
(Ⅰ)取360μl自然全血，需注意在2分钟内完成检测。
(Ⅱ)用带有抗凝剂的枸橼酸钠采血管取血。
在sonoclot仪器上运行：
(Ⅰ)全血样本：将全血样本从注射器转移到试剂杯中，转移容量为360μL，点击Start开始混匀，待混合结束时显示屏出现Close Head字样时关闭分析仪头。
(Ⅱ)枸橼酸钠抗凝血：上下轻轻颠倒采血管，取500μL加入1.5ML离心管中，再取20μL0.25M氯化钙加入到1.5ML离心管中上下颠倒3次从中用移液器吸360μL放入准备好的试剂杯里面。点击Start开始混匀，待混合结束时显示屏出现Close Head字样时关闭分析仪头。
(Ⅲ)开始之后数分钟。样本开始变为凝血块，Sonoclot分析仪会检测到最初凝血块生成，发出哔声，并显示激活凝血时间(ACT)。再接着数分钟内，纤维蛋白原开始转化成纤维蛋白单体，凝血信号开始增加，分析仪会根据凝血信号计算出凝血速率(CR)。紧接着数分钟内，分析仪继续监测血液样本的粘弹性改变，以量化血小板活化和血块凝缩，以此监测血小板功能(PF)。
本发明试剂与进口试剂SIENCO公司的gbACT+试剂盒的比较，gbACT+试剂盒是临床上应用最广泛的凝血和血小板功能检测的试剂盒，也是同类产品口碑最好。
所述试剂与gbACT+比较，所述试剂配方和gbACT+每一种准备6个，2个临床随机取用血液样本，在Sonoclot凝血和血小板功能分析仪型号SCP2上测试比较2个配方。
所述试剂与gbACT+比较结果见表一：
表一
Claims (9)
1.一种检测凝血和血小板功能的试剂，其特征在于，所述试剂是玻璃珠，所述玻璃珠粒径分布在0.1-60μm，单个玻璃珠的重量为0.001-0.01g。
2.根据权利要求1所述的检测凝血和血小板功能的试剂，其特征在于，所述玻璃珠是球形玻璃珠。
3.根据权利要求1所述的检测凝血和血小板功能的试剂，其特征在于，所述玻璃珠中二氧化硅的含量>90％。
4.根据权利要求1所述的检测凝血和血小板功能的试剂，其特征在于，所述玻璃珠的D50为15-25μm。
5.一种试剂盒，其特征在于，包括权利要求1-3任一项所述的检测凝血和血小板功能的试剂。
6.权利要求1-4任一项所述的检测凝血和血小板功能的试剂、或权利要求4所述的试剂盒在添加肝素血液样本的枸橼酸钠抗凝全血检测中的应用。
7.根据权利要求6所述的应用，其特征在于，所述添加肝素，是每毫升添加0-2单位。
8.权利要求1-4任一项所述的检测凝血和血小板功能的试剂、或权利要求4所述的试剂盒在自然全血样本检测中的应用。
9.权利要求1-4任一项所述的检测凝血和血小板功能的试剂、或权利要求4所述的试剂盒在Sonoclot检测仪检测中的应用。
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