CN1962876A - 酶法测定d－3－羟基丁酸试剂盒 - Google Patents

酶法测定d－3－羟基丁酸试剂盒 Download PDF

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CN1962876A
CN1962876A CN 200610151021 CN200610151021A CN1962876A CN 1962876 A CN1962876 A CN 1962876A CN 200610151021 CN200610151021 CN 200610151021 CN 200610151021 A CN200610151021 A CN 200610151021A CN 1962876 A CN1962876 A CN 1962876A
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test kit
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于永光
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Abstract

本发明涉及一种酶法测定D－3－羟基丁酸试剂盒。本产品是生物工程领域高科技产品，是一种体外诊断试剂。本产品经检索填补了国内空白，本产品经多年研究，解决了酶试剂盒的稳定性问题，省略了原来试剂盒需要冷冻干燥的工艺，降低成本，使用者更加方便。为解决这些问题试剂盒使用了新型防腐剂2－甲基－4－异噻唑－3－酮、抗干扰试剂吐温－80、新型缓冲剂3羟甲基氨基甲烷－盐酸缓冲系统，酶稳定剂乳糖。这种配方可以使试剂盒由冻干型改为液体型，本产品主要提供全国大中小医院检验科使用，糖尿病酮症酸中毒是一临床常见病多发病，原因是血酮体增高引起，D－3－羟基丁酸是酮体重要组成部分，占酮体总量70％，酮症诊断治疗不及时将有生命危险，D－3－羟基丁酸检测对糖尿病酮症酸中毒具有特异的诊断价值。

Description

酶法测定D-3-羟基丁酸试剂盒
技术领域；本发明涉及一种酶法测定D-3-羟基丁酸试剂盒。该产品为生物工程领域高科技产品，是国家按医疗器械管理的体外诊断试剂。
背景技术；糖尿病酮症酸中毒是一临床常见病，多发病，原因是血酮体增高引起，D-3-羟基丁酸酮体重要组成部分，占酮体总量70％，酮症诊断治疗不及时将有生命危险，D-3-羟基丁酸测定有显色法，气相色谱法，放射化学法，毛细管等速电泳法等。放射化学法因有射线，化学品难于接受，毛细管等速电泳操作时间长，复杂不适于批量检测。气相色谱特导性差。目前，在《临床化学诊断方法大全》中介绍了酶法测定D-3-羟基丁酸方法，但未涉及D-3-羟基丁酸试剂盒，文献介绍的方法只能现配现用，不能长期保存，本试剂盒可冷藏保存一年以上，长期使用。
发明内容；本发明的目的在于提供一种长期保存的酶法D-3-羟基丁酸测定试剂盒。
1本发明的技术方案：一种酶法测定D-3-羟基丁酸试剂盒。其特征在于试剂盒含有D-3-羟基丁酸脱氢酶、氧化型辅酶I、3羟甲基氨基甲烷、吐温-80、2-甲基-4-异噻唑-3-酮、乳糖。各组份含量最佳毫摩尔比为每升3羟甲基氨基甲烷60-140，氧化型辅酶I 2.0-3.4，D-3-羟丁酸脱氢酶30-100IU，保护剂兼赋型剂乳糖1-5克，2-甲基-4-异噻唑-3-酮0.1-0.6活性单位、吐温-80 0.1-0.6克。上述成分加水至一升，调PH在6.5-7.1
2.意义陈述；本发明配方中，3羟甲基氨基甲烷作为缓冲剂有维持PH的作用，吐温-80有乳化作用，D-3-羟基丁酸脱氢酶起催化剂作用、2-甲基-4-异噻唑-3-酮起防腐作用，保护剂乳糖对D-3-羟基丁酸脱氢酶有保护作用、氧化型辅酶I参与底物反应，以上各成分含量浓度为测量D-3-羟基丁酸最佳浓度值。
本发明的有益效果：本发明配方中3羟甲基氨基甲烷作为缓冲剂有维持PH的作用，吐温-80有乳化作用，D-3-羟基丁酸脱氢酶起催化剂作用、2-甲基-4-异噻唑-3-酮起防腐作用，保护剂乳糖既对β-羟丁酸脱氢酶有保护作用、氧化型辅酶I参与底物反应，试剂抗干扰能力强，中等程度溶血，乳糜血，血糖20mmOl/l以下，胆红素100umol/l以下对测定不产生干扰，本试剂盒为液体试剂，有效期一年。冷藏一年试剂盒中酶活力损失小于5％。
具体实施方式；取超纯水一升用前将水加热至沸，密封备用。将3羟甲基氨基甲烷60-140mmol，氧化型辅酶I 2.0-3.4mmOl，D-3-羟基丁酸脱氢酶30-100IU，保护剂乳糖1-5克，2-甲基-4-异噻唑-3-酮0.1-0.6活性单位、吐温-80 0.1-0.6克。上述成分加水至一升，调PH在6.5-7.1，将试剂溶液分装成5-10毫升/瓶，随时备用。本试剂盒比原来试剂盒的优越性是省去了冷冻干燥工艺，降低成本，使用方便。

Claims (1)

  1. 本发明涉及一种酶法测定D-3-羟基丁酸试剂盒。
    1.一种酶法测定D-3-羟基丁酸试剂盒，其特征在于含有D-3-羟基丁酸脱氢酶、氧化型辅酶I、3羟甲基氨基甲烷、吐温-80、2-甲基-4-异噻唑-3-酮、乳糖。各组份含量最佳毫摩尔比为每升3羟甲基氨基甲烷60-140，氧化型辅酶I 2.0-3.4，D-3-羟基丁酸脱氢酶30-100IU，保护剂乳糖1-5克，2-甲基-4-异噻唑-3-酮0.1-0.6活性单位、吐温-80 0.1-0.6克。
    意义陈述
    本发明配方中本发明配方中3羟甲基氨基甲烷作为缓冲剂有维持PH的作用，吐温-80有乳化作用，D-3-羟基丁酸脱氢酶起催化剂作用、2-甲基-4-异噻唑-3-酮起防腐作用，保护剂乳糖对D-3-羟基丁酸脱氢酶有保护作用、氧化型辅酶I参与底物反应，试剂抗干扰能力强，中等程度溶血，乳糜血，血糖20mmOl/l以下，胆红素100umol/l以下对测定不产生干扰，本试剂盒为液体试剂，有效期一年。冷藏一年试剂盒中酶活力破坏小于5％，氧化型辅酶I小于10％，本试剂盒线性范围达5.0mmol/l，正常参考范围0.038-0.286mmol/l，回收率96.5-103.8％之间，变异系数CV＝5.7％，X＝0.26mmol/l，SD＝0.015，n＝20。由3羟甲基氨基甲烷组成的缓冲系统作为缓冲剂PH6.8以上各成分含量浓度为测量D-3-羟基丁酸最佳浓度值。
    本发明的有益效果：
    3羟甲基氨基甲烷作为缓冲剂有维持PH的作用，吐温-80有乳化作用，D-3-羟基丁酸脱氢酶起催化剂作用、2-甲基-4-异噻唑-3-酮可防腐，本试剂盒使用的保护剂可有效稳定氧化型辅酶I。试剂抗干扰能力强，中等程度溶血，乳糜血，血糖20mmOl/l以下，胆红素100umol/l以下对测定不产生干扰，冻干试剂冷藏一年，试剂盒中活力小于5％，还原型辅酶I破坏小于10％，本试剂盒线性范围达5.0mmol/l，正常参考范围0.038-0.286mmol/l，回收率96.5-103.8％之间，变异系数CV＝5.7％，X＝0.26mmol/l，SD＝0.015，n＝20。本项产品为液体试剂无需冷冻干燥，冷藏有效期一年。
CN 200610151021 2006-11-13 2006-11-13 酶法测定d－3－羟基丁酸试剂盒 Pending CN1962876A (zh)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102405294A (zh) * 2009-04-28 2012-04-04 布尔曼实验室股份公司 适用于测定样品中γ-羟基丁酸(GHB)浓度的方法、组合物和药盒
CN105510261A (zh) * 2015-11-30 2016-04-20 山东博科生物产业有限公司 一种灵敏度高的d3-羟丁酸（d3h）检测试剂及其检测方法
CN108350439A (zh) * 2015-12-21 2018-07-31 豪夫迈·罗氏有限公司 来自粪产碱菌的突变体3-羟基丁酸脱氢酶及其相关方法和用途

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102405294A (zh) * 2009-04-28 2012-04-04 布尔曼实验室股份公司 适用于测定样品中γ-羟基丁酸(GHB)浓度的方法、组合物和药盒
CN105510261A (zh) * 2015-11-30 2016-04-20 山东博科生物产业有限公司 一种灵敏度高的d3-羟丁酸（d3h）检测试剂及其检测方法
CN108350439A (zh) * 2015-12-21 2018-07-31 豪夫迈·罗氏有限公司 来自粪产碱菌的突变体3-羟基丁酸脱氢酶及其相关方法和用途

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