CN109297919A - 一种抗肝素干扰的c反应蛋白检测试剂盒 - Google Patents
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Abstract
本发明涉及临床体外检测试剂技术领域，特别涉及一种抗肝素干扰的C反应蛋白检测试剂盒。本发明试剂盒是双试剂液体型试剂盒，试剂R1包括乙二胺四乙酸二钠、磷酸氢二钠、磷酸二氢钾、氯化钠、聚乙二醇，试剂R2包括乙二胺四乙酸二钠、磷酸氢二钠、磷酸二氢钾、氯化钠、聚乙二醇、CRP抗体。本发明试剂组分中聚乙二醇6000投料减少，可解决肝素干扰问题，具有很好的市场潜力。
Description
技术领域
本发明涉及临床体外检测试剂技术领域，特别涉及到血清或血浆中的C反应蛋白含量的检测。
背景技术
C反应蛋白是一种急性时相反应蛋白，在机体发炎时，血清中的CRP升高。C反应蛋白水平增加常作为风湿症活动期的指标或用于提示手术中组织损伤的程度和术后炎症与细菌感染的鉴别，C反应蛋白水平增加亦可见于慢性类风湿性关节炎、全身性红斑狼疮、慢性进行性肝炎、肝硬化、心肌损伤、恶性肿瘤(特别是全身性转移)等。
本发明使用的C反应蛋白检测诊断方法是免疫比浊法。
前期实验R1中聚乙二醇6000按照60克/升投料，配好试剂后用肝素处理后的样本测试曲线R1段上升导致结果异常。
鉴于以上因素，发明了一种抗肝素的C反应蛋白检测试剂盒。该方法具有延长试剂稳定、抗肝素干扰等优势，从而为CRP检测方法学提供新的资料。
发明内容
本发明内容在于解决CRP检测试剂盒不抗肝素的现象。
基本原理：本试剂采用免疫比浊法测定人血清中C反应蛋白含量。在氨基乙酸缓冲体系下，样本中的CRP与CRP抗体试剂反应，形成免疫复合物，在波长340nm处引起吸光度的改变，检测其浊度变化，其变化程度与样本中的CRP含量成正比。
应用本发明试剂盒测定血清中CRP含量方法如下：检测波长为340nm，样本量：试剂R1：试剂R2＝13ul：180ul：60ul。
本发明是一种抗肝素干扰的CRP检测试剂盒，本发明试剂盒是由试剂R1和试剂R2两种试剂组成的液体型双试剂检测诊断试剂，其中试剂R1组成为：
试剂R2包含：
本发明试剂盒一种抗肝素干扰的CRP检测试剂盒，试剂R1和R2各物质的选择依据是：
乙二胺四乙酸二钠是一种离子螯合剂，它能有效的螯合金属离子，防止一些杂质离子对检测反应引起不必要的干扰；
磷酸盐缓冲液主要是缓冲作用，保证反应时外环境的稳定，致使检测结果不会受到波动和影响；
氯化钠用于模拟人体内血液环境，保证检测的真实性和可靠性；
聚乙二醇可起到抗干肝素干扰的作用。
本发明可以应用到迪瑞、迈瑞、日立、东芝等系列的全自动化分析仪中进行自动化检测。
应用本发明进行体外样本诊断检测时，操作方法如下，设置波长为340nm，终点法，样本量：试剂R1：试剂R2＝13ul：180ul：60ul。样本针吸取样本后于试剂R1中进行孵育，以保证反应环境外界条件的稳定，然后读取吸光光度值为ΔA1，进而加入试剂R2，样本中的CRP与CRP抗体试剂反应，形成免疫复合物，在波长340nm处引起吸光度的改变，检测其浊度变化，其变化程度与样本中的CRP含量成正比。
37℃反应5-10分钟左右读取吸光光度值为ΔA2，通过计算吸光光度值的变化，进而计算出CRP的含量。计算公式如下
CRP含量＝标准液浓度*吸光光度值(ΔA2-ΔA1)
相对于市场上目前流通的免疫比浊法CRP检测试剂盒，本发明试剂盒抗肝素干扰，适用于全自动生化分析仪，临床使用价值较大。
具体实施方式
以下结合实施例对本发明进行进一步说明。
实施例1
没改进之前的方案，所述的试剂R1为：
试剂R2：
实施例2
根据本发明配置如下试剂，所述的试剂R1为：
试剂R2包含：
抗肝素性能验证：
运用实施例1、实施例2试剂和目前市场上流通的试剂盒进行对比，使用仪器为全自动生化分析仪BS-800，参数设置如下，波长为340nm，反应方向为正方向，读数点设置为14-16点和31-33点，样本量：试剂R1：试剂R2＝13ul：180ul：60ul。检测结果如表1所示：
表1实施例1、2与对照试剂检测肝素样本结果对比
从数据看，我们发现本发明测试肝素样本的准确度明显优于改进前的，为本发明试剂盒提供了良好的发展空间，同时增强了本发明试剂盒在市场上竞争力。
Claims (3)
1.一种抗肝素干扰的C反应蛋白检测试剂盒，其特征在于，本发明试剂盒是双试剂液体型试剂盒，具体组成成分为：
试剂R1：
乙二胺四乙酸二钠 0.5%
磷酸氢二钠 1.2%
磷酸二氢钾 0.5%
氯化钠 0.87%
聚乙二醇6000 40g/l
试剂R2：
乙二胺四乙酸二钠 0.5%
磷酸氢二钠 1.2%
磷酸二氢钾 0.5%
氯化钠 0.87%
聚乙二醇6000 2.5%
CRP抗体 20%。
2.根据权利要求1所述的试剂盒，其特征在于本发明试剂盒试剂Ｒ1和试剂Ｒ２的使用比例为3:1。
3.根据权利要求1所述的试剂盒，其特征在于试剂R1所用到的原料聚乙二醇6000投料减少，用此可解决肝素干扰问题。
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