CN110082346A - 一种高脂血处理试剂及其在载脂蛋白e检测中的应用 - Google Patents

一种高脂血处理试剂及其在载脂蛋白e检测中的应用 Download PDF

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CN110082346A
CN110082346A CN201910391404.0A CN201910391404A CN110082346A CN 110082346 A CN110082346 A CN 110082346A CN 201910391404 A CN201910391404 A CN 201910391404A CN 110082346 A CN110082346 A CN 110082346A
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reagent
perfluoropolyether
detection
apo
upper layer
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CN110082346B (zh
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华权高
沈鹤霄
徐春雷
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Wuhan Life Origin Biotech Joint Stock Co Ltd
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Abstract

一种去除高血脂血清中脂类的试剂及其在载脂蛋白E检测过程中的应用,包括如下步骤：将血清和去除血脂的试剂混合，搅拌混匀后静置10分钟，血清出现分层，下层为无血脂的血清，上层白色的为分离出来的血脂部分。

Description

一种高脂血处理试剂及其在载脂蛋白E检测中的应用
技术领域
本发明涉及医学检测领域，具体涉及一种高脂血处理试剂及其在血清载脂蛋白E检测中的应用。
背景技术
脂血为临床生化项目检测的常见干扰现象，其血浆呈浑浊状，若乳糜微粒含量过多，血浆可呈乳白色，所以也称“乳糜血”。由于乳糜微粒颗粒较大，可对入射光产生散射，导致某些生化结果出现偏差。近年来，不少研究者采用真空超速离心、高速离心、PEG沉淀或乙醚处理等方法来消除脂血对常规生化项目的干扰，但各有优缺点。LipoClear不适合用于从特定样品中去除脂血后进行葡萄糖，钠，钾，氯化物，磷酸盐，镁，CK-MB， ALP，GGT，总蛋白，白蛋白，CRP和肌钙蛋白T测量。
载脂蛋白E主要存在于CM、VLDL、IDL和部分HDL中。ApoE的浓度与血浆甘油三酯含量呈正相关。ApoE是LDL受体的配体，也是肝细胞CM残粒受体的配体，它与脂蛋白代谢密切相关；它具有多态性，多态性与个体血脂水平及动脉粥样硬化发生发展密切相关，参与激活水解脂肪的酶类，参与免疫调节及神经组织的再生。
发明内容
本发明的目的是为了解决上述问题而提供。
本发明通过以下技术方案实现：
在高血脂血清样本中添加适量全氟聚醚液体，经过全氟聚醚处理后分成上下两层，上层为分离的血脂成分，下层为澄清的血清部分。具体操作为取中高浓度脂血血清0.5mL于小型离心管中,按试剂说明书加入0.1mL 全氟聚醚试剂，震荡混均后静置5min，最后采用6000r/min离心5min，使用加样枪吸取上层凝固物下的清亮液体约0.3mL，测定载脂蛋白E项目。载脂蛋白E的试剂盒组分如下：
试剂1的组分是R1：
R2：检测试剂2
磷酸盐缓冲液 100mmol/L
羊抗人ApoE抗体 14.2ml/L
使用该试剂盒的方法为：
1.基本参数：
方法：终点法 温度：37℃
主波长：340nm 副波长：700nm
标本量：3μl R1：225μl
R2：75μl 反应方向：正向
反应时间：10min 定标方式：多点定标
2.操作：
混匀，37℃恒温5分钟测定各管吸光度A2。
计算ΔA＝A2-A1。
3.计算：
根据多点校准品浓度和对应吸光度变化值ΔA，采用非线性校准法确定工作曲线，所测样本吸光度变化在工作曲线上相对应的浓度值即为所测浓度。
附图说明
图1高血脂血清处理前
图2高血脂血清处理后
具体实施方式
下面结合附图和实施方式对本发明做进一步的详细说明。以下实施例仅是范例性的，仅用以对本发明的技术方案做更进一步的详细说明，本领域的普通技术人员应当理解，在不偏离本发明技术方案的精神和范围内，对技术方案进行的修改或者替换均应涵盖在本发明的权利要求保护范围内。
实施例1载脂蛋白E检测试剂盒的制备
试剂1的组分是R1：
R2：检测试剂2
磷酸盐缓冲液 100mmol/L
羊抗人ApoE抗体 14.2ml/L
实施例2去脂剂的对比实验
在高血脂血清样本中添加适量全氟聚醚液体，经过全氟聚醚处理后分成上下两层，上层为分离的血脂成分，下层为澄清的血清部分。先测定ApoE 项目，再按试剂说明书加入0.1mL试剂，震荡混均后静置5min，最后采用 6000rpm离心5min，使用加样枪吸取表层凝固物下的清亮液体约0.3mL，再次测定上述ApoE项目。检测数据根据稀释倍数进行校正。统计学处理采用SPSS17.0软件，处理前后ApoE指标的平均偏差(Bias％)＜1/2美国临床医学检验部门修正法规(CLIA′88)允许总误差(TEa)为临床可接受水平。
实验例1对照组原始标本检测结果与加入人工脂肪乳，经脂质清除剂处理后ApoE指标的结果见表1，结果显示ApoE项目处理前后，该项目的平均偏差(Bias％)＜1/2TEa，属临床可接受水平，其误差是由随机误差引起。
表1 40例对照组标本处理前后项目检测结果
实验例2.试剂盒检测参数测定：
测定范围：5mg/L-100mg/L，r≥0.990。
精密度：批内CV≤4.7％、批间相对极差≤7.3％。
准确度：不准确度≤10％。
空白吸光度：本试剂在340nm波长处，10mm光径下，吸光度值≤0.100。

Claims (4)

1.一种高脂血处理试剂及其在载脂蛋白E检测中的应用，其特征在于，在高血脂血清样本中添加适量全氟聚醚液体，经过全氟聚醚处理后分成上下两层，上层为分离的血脂成分，下层为澄清的血清部分。
2.如权利要求1所述全氟聚醚使用方法，其特征在于：取中高浓度脂血血清0.5mL于小型离心管中,按试剂说明书加入0.1mL全氟聚醚试剂，震荡混均后静置5min，最后采用6000r/min离心5min，使用加样枪吸取上层凝固物下的清亮液体约0.3mL，测定载脂蛋白E项目。
3.一种载脂蛋白E检测试剂盒，包括试剂1，试剂2，标准品，其特征在于：在试剂盒检测乳白色高脂血清之前加入如权利要求1所述的高血脂处理试剂，试剂盒中的R1含有去脂剂全氟聚醚，其中试剂1的组分是R1：
磷酸盐缓冲液 100mmol/L
PEG 31g/L
全氟聚醚 0.5mL
防腐剂 0.03%
R2：检测试剂2
磷酸盐缓冲液 100mmol/L
羊抗人ApoE抗体 14.2ml/L。
4.所述防腐剂为叠氮钠，Proclin300，硫柳汞等。
CN201910391404.0A 2019-05-12 2019-05-12 一种载脂蛋白e检测试剂盒 Active CN110082346B (zh)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103415349A (zh) * 2011-11-08 2013-11-27 瑞尔赛特股份有限公司 使用辅助液体分离悬浮液中各组分的方法与系统
CN104722342A (zh) * 2009-03-24 2015-06-24 芝加哥大学 滑动式芯片装置和方法
US20160326378A1 (en) * 2015-05-06 2016-11-10 Korea Institute Of Ocean Science & Technology Amphiphilic hydrogel particles for antifouling paint and method of fabricating the same
JP2017047340A (ja) * 2015-08-31 2017-03-09 ジャパン・フィールド株式会社 被洗浄物の蒸気洗浄方法及びその装置

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104722342A (zh) * 2009-03-24 2015-06-24 芝加哥大学 滑动式芯片装置和方法
CN103415349A (zh) * 2011-11-08 2013-11-27 瑞尔赛特股份有限公司 使用辅助液体分离悬浮液中各组分的方法与系统
US20160326378A1 (en) * 2015-05-06 2016-11-10 Korea Institute Of Ocean Science & Technology Amphiphilic hydrogel particles for antifouling paint and method of fabricating the same
JP2017047340A (ja) * 2015-08-31 2017-03-09 ジャパン・フィールド株式会社 被洗浄物の蒸気洗浄方法及びその装置

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