本发明专利技术涉及血清钾检测技术领域，特别涉及一种血清钾检测试剂，试剂R1中含有缓冲液、穴合剂，PEP，ADP，甲酸，甲酸脱氢酶，α-酮戊二酸，NADH，GLDH（谷氨酸脱氢酶），聚乙二醇6000，乙二醇，甘露醇，海藻糖，BSA，PK（丙酮酸激酶），烷基糖苷（APG），防腐剂；试剂R2中含有缓冲液，聚乙二醇6000，乙二醇，甘露醇，海藻糖，BSA，LDH（乳酸脱氢酶），烷基糖苷（APG），防腐剂。采用Tris缓冲液，添加多种稳定剂，且引入的NADH循环再生系统，显著改善了试剂的稳定性；并且添加的新型非离子型表面活性剂烷基糖苷（APG），可有效防止反应体系浑浊，显著提高了试剂的灵敏度和稳定性。
【技术实现步骤摘要】
本专利技术涉及血清钾检测
，特别涉及一种血清钾检测试剂，还涉及使用此 检测试剂的检测方法。
技术介绍
钾（potassium，K)是人体内维持细胞生理活动的主要阳离子，在保持机体的正 常渗透压及酸碱平衡、参与糖及蛋白代谢、保证神经肌肉的正常功能等方面具有重要作用。 钾离子大部分（98%)存在于细胞内，少量存在于细胞外液，且浓度恒定。正常人血清中含K+ 约4~5mmol/L。体内的钾离子经常不断地在细胞内与体液之间相互交换，以保持动态平 衡。血清钾、钠、氯测定是临床常见的组合检测项目之一，有助于水、电解质平衡和酸碱平衡 紊乱的判断。 目前，钾离子检测方法主要有离子选择电极法（ISE)、火焰光度法和酶法。其中 离子选择电极法（ISE)和酶法在临床上最为常用。ISE法具有标本用量少，可与自动生化 分析仪组合，快速准确，重复性好等优点。但该法电极具有一定寿命，需要定期更换，费用较 高。而酶法的精密度和准确度可以与火焰光度法接近，且具有简便、精确、重复性好、特异性 强、血清用量少等优点，但存在稳定性较差，易受干扰等缺点。 鉴于此，本专利技术在酶法的基础上优化反应体系，添加稳定剂聚乙二醇6000、甘露 醇、海藻糖、BSA、乙二醇等可有效提高试剂的稳定性；并且通过引入辅酶NADH再生系统，可 以有效降低NADH的氧化，大大增强试剂稳定性。优选的新型非离子表面活性剂烷基糖苷 (APG)的加入可防止反应体系浑浊，增强底物的稳定性，提高试剂的抗干扰能力。该试剂操 作简便快速，适用于自动化分析，是一种更加稳定、抗干扰能力强的血清钾（K)试剂。【
技术实现思路
】 本专利技术的目的是提供一种用于检测血清钾（K)的试剂及使用该试剂检测血清钾 含量的方法。该试剂盒采用酶法，可以有效检测血清钾的含量，抗干扰能力强，稳定性好等 优点。 基本原理： 通过钾依赖性丙酮酸激酶催化底物磷酸烯醇式丙酮酸（PEP)的酶动力学反应检测钾， 其产物丙酮酸盐在乳酸脱氢酶（LDH)作用下与NADH反应生成NAD+，其在340nm的吸光值 下降与钾浓度呈比例。并且，本试剂引入辅酶NADH再生系统，通过添加少量的的甲酸脱氢酶，可以在不干扰 主体反应的前提下，将氧化态NAD+转化还原态NADH，使反应底物NADH在试剂存储过程中可 以始终保持稳定，显著提高了试剂的稳定性。本专利技术是通过以下步骤得到的： 一种血清钾检测试剂，包括试剂R1和试剂R2,所述试剂R1和试剂R2的组成如下： 试剂R1中含有 缓冲液........................................200mmol/L, 穴合齐U........................................14mmol/L, PEP...........................................4mmol/L, ADP...........................................3. 5mmol/L, 甲酉爱..........................................0. 2mmol/L, 甲酸脱氢酶.....................................100U/L， α-酮戊二酸....................................3mmol/L， NADH..........................................0· 35mmol/L， GLDH(谷氨酸脱氢酶）..............................12KU/L， 聚乙二醇 6000...................................10g/L， 乙二醇.........................................5ml/L， 甘露醇.........................................20g/L， 海藻糖.........................................l〇g/L, BSA............................................3g/L， PK(丙酮酸激酶）..................................2KU/L， 烷基糖苷（APG)...................................lg/L， 防腐齐U.........................................lml/L； 2)试剂R2的组分为： 缓冲液.........................................20mmol/L， 聚乙二醇 6000...................................10g/L， 乙二醇.........................................5ml/L， 甘露醇.........................................20g/L， 海藻糖.........................................l〇g/L, BSA.............................................3g/L， LDH(乳酸脱氢酶）..................................65KU/L， 烷基糖苷（APG)....................................lg/L， 防腐齐U..........................................lml/L。 所述的血清钾检测试剂，试剂R1中缓冲液为25°C，pH为8. 2的Tris缓冲液。 所述的血清钾检测试剂，试剂R2中缓冲液为25°C，pH为9. 0的Tris缓冲液。 所述的血清钾检测试剂，所述防腐剂为Proclin300。 所述的血清钾检测试剂来检测血清钾含量的检测方法，使用全自动生化分析仪利 用固定时间法进行测定，检测主波长为340nm。 所述的检测方法，R1试剂和R2试剂的比例为3:1。 本专利技术的有益效果： 1) 优化反应体系并添加聚乙二醇6000、甘露醇、海藻糖、BSA、乙二醇等多种稳定剂，可 以显著改善了试剂的稳定性和抗干扰能力； 2) 采用新型非离子表面活性剂烷基糖苷（APG)的加入可防止反应体系浑浊，增强底物 的稳定性，提高试剂的抗干扰能力。 3)引入辅酶NADH再生系统，通过添加少量的的甲酸脱氢酶，可以在不干扰主体反 应的前提下，将氧化态NAD+转化还原态NADH，使反应底物NADH在试剂存储过程中可以始终 保持稳定，显著提高了试剂的稳定性。 4)试剂的准确度和稳定性良好，价格便宜，使用方便，完全可以满足临床需要。【附图说明】 图1为两种试剂的相关性曲线图， 图2为两种试剂效期稳定性曲线图。【具体实施方式】 下面结合具体实施例对本专利技术进行进一步说明： 实施例1 血清钾的检测试剂，包试剂R1和试剂R2 : 1)其R1的组成为： Tri本文档来自技高网...
【技术保护点】
一种血清钾检测试剂，其特征在于包括试剂R1和试剂R2，所述试剂R1和试剂R2的组成如下：试剂R1中含有：缓冲液···········································································200mmol/L，穴合剂········································································14mmol/L，PEP···············································································4mmol/L，ADP··············································································3.5mmol/L，甲酸·············································································0.2mmol/L，甲酸脱氢酶···································································100U/L，α‑酮戊二酸·································································3mmol/L，NADH·············································································0.35mmol/L，GLDH（谷氨酸脱氢酶）···················································12KU/L，聚乙二醇6000································································10g/L，乙二醇··············································································5ml/L，甘露醇··············································································20g/L，海藻糖·················································································10g/L，BSA·······················································································3g/L，PK（丙酮酸激酶）································································2KU/L，烷基糖苷（APG）································································1g/L，防腐剂·················································································1ml/L；试剂R2的组分为：缓冲液···········································································20mmol/L，聚乙二醇6000································································10g/L，乙二醇··············································································5ml/L，甘露醇··············································································20g/L，海藻糖·················································································10g/L，BSA·····························································...
【技术特征摘要】
【专利技术属性】
技术研发人员：李志明，谭柏清，李静，
申请(专利权)人：山东博科生物产业有限公司，
类型：发明
国别省市：山东;37
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