CN113584124A - 复合稳定剂及含其的鸟嘌呤脱氨酶测定试剂盒 - Google Patents
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Abstract
本发明公开了一种复合稳定剂及含其的鸟嘌呤脱氨酶测定试剂盒,所述复合稳定剂由各物质按以下质量浓度组成:庆大霉素0.002g/L~1.0g/L、牛血清白蛋白(BSA)0.1g/L~10g/L、甘露醇10g/L~100g/L;在鸟嘌呤脱氨酶测定试剂盒中添加所述复合稳定剂,能有效延长鸟嘌呤脱氨酶测定试剂盒的保存时间,有利于该试剂盒在市场中的进一步推广。
Description
技术领域
本发明属于临床检验生化分析技术领域,特别是涉及一种复合稳定剂及含其的鸟嘌呤脱氨酶测定试剂盒。
背景技术
鸟嘌呤脱氨酶(GUA)主要定位于肝脏,在骨骼肌、心肌、胰腺、肺等几乎缺如,其肝脏特异性远大于谷丙转氨酶(ALT)和天门冬氨酸氨基转移酶(AST),在急性肝炎早期诊断及鉴别诊断、输血后肝炎的预防等方面已经引起重视,尽管鸟嘌呤脱氨酶活性检测对肝病的诊断及预防具有重大意义,但到目前为止,仍没有令人满意的即稳定又灵敏的GUA全自动分析用试剂盒。
当前,常使用以下方法对GUA活性进行测定:1、Rousca&Norris.E.R;Arch.Biochem法,其利用鸟嘌呤与黄嘌呤的吸光度不同,测定鸟嘌呤的减少量,进而计算GUA的活性;2、测量尿酸法,该方法中样本内含的GUA作用于底物鸟嘌呤生成黄嘌呤,黄嘌呤在黄嘌呤氧化酶(XOD)的作用下生成尿酸和过氧化氢,尿酸在290nm处有吸收峰,通过检测尿酸的生成量计算GUA的活性;3、氨定量法,使用GUA作用于底物鸟嘌呤生成黄嘌呤和氨,利用谷氨酸脱氢酶法测定NADH的生成量进而计算GUA的活性;4、过氧化氢定量法,包括eintz Fr1tz、ReckelSylvia.&.Kalden Joachim、R、Enzyme法和杉浦法,均通过测量过氧化氢的生成量来定量计算GUA的活性;5、14C标记法,使用14C标记的鸟嘌呤生成黄嘌呤,通过电泳分离,并用闪烁计数器测量其放射性,计算GUA的活性;然而上述方法1-3中消光系数小、灵敏度低,而14C标记法需使用特殊设备,不适用于临床,杉浦法同样存在反应时间长,内源性干扰大的问题。
在实现本发明过程中,发明人发现现有技术中至少存在以下问题:目前临床检查用的鸟嘌呤脱氨酶测定试剂盒多以液体或冻干的形式供应,在测定过程中液体以原样使用,冻干产品溶解后使用,测定后剩余试剂在打开状态于2-8℃的环境中保存,而GUA测定试剂盒测量值和灵敏度会随储存时间的推移而降低,GUA测定试剂盒在存储期间的稳定性不能令人满意,现有技术对冻干测定试剂盒具有稳定作用,但在液体测定试剂盒中未必有效,需开发更为有效的稳定方法。
发明内容
本发明实施例的目的在于提供一种复合稳定剂及含其的鸟嘌呤脱氨酶测定试剂盒,使用所述复合稳定剂能保护液态GUA测定试剂盒中的工具酶不失活,以此延长鸟嘌呤脱氨酶测定试剂的保存时间,有利于其在市场上进一步推广。
本发明实施例所采用的技术方案是,复合稳定剂,由各组分按以下质量浓度组成:庆大霉素0.002g/L~1.0g/L、牛血清白蛋白0.1g/L~10g/L、甘露醇10g/L~100g/L。
进一步的,由各组分按以下质量浓度组成:庆大霉素0.1g/L~0.5g/L、牛血清白蛋白1g/L~8g/L、甘露醇20g/L~80g/L。
进一步的,由各组分按以下质量浓度组成:庆大霉素0.2g/L~0.3g/L、牛血清白蛋白2g/L~5g/L、甘露醇30g/L~50g/L。
鸟嘌呤脱氨酶测定试剂盒包含第一试剂、第二试剂和复合稳定剂,其中第一试剂组成为:0.1M Tris Buffer pH7.6,鸟嘌呤盐酸盐0.1M,尿酸酶40U/L,过氧化氢酶17KU/L,4-氨基安替比林0.05mM;
第二试剂组成为:0.1M Tris Buffer pH6.5,黄嘌呤氧化酶(XOD)240U/L,过氧化物酶(POD)8KU/L,超氧化物歧化酶(SOD)24KU/L,TOOS 0.1M,叠氮钠3.2mM。
进一步的,所述第一试剂与第二试剂的体积比为4:1。
本发明实施例的有益效果是:复合稳定剂中包含的庆大霉素能够保护XOD的酶蛋白质结构,防止酶蛋白质表面构象发生变化,抑制XOD随时间推移产生的活性降低,延长鸟嘌呤脱氨酶活性测定试剂盒的稳定期;牛血清白蛋白和甘露醇同样具有保护酶、稳定酶的胶体作用,及保护酶蛋白的立体构象作用。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1是本发明实施例的效果图。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
GUA测定试剂盒由多种酶、色原物质、电子传递物质及缓冲物质组成的复合物,在该反应体系中,任一成分发生改变,特别是试剂中工具酶的活性发生变化,都会影响整个试剂盒的质量,使其劣化,本发明实施例使用工具酶再次添加法,在发生劣化的GUA测定试剂盒中分别加入不同的工具酶,逐项排除各项工具酶对检测结果的影响,具体实验结果如表1所示:
表1劣化GUA测定试剂盒加酶后的测定结果
通过对GUA测定试剂盒的劣化机理进行研究,发现当加入所需浓度的XOD时,GUA测定试剂盒的测定结果恢复初始值,即GUA测定试剂盒劣化的主要原因是检测试剂中的黄嘌呤氧化酶(XOD)的残存活性随保存时间的延长而降低,导致黄嘌呤被氧化成尿酸和过氧化氢的速度减慢,GUA测定试剂盒的灵敏度下降,不能准确检测鸟嘌呤脱氨酶的含量。
本发明通过在GUA测定试剂盒中添加复合稳定剂,以保持XOD的立体结构,使GUA测定试剂盒的长期稳定性良好,GUA测定试剂盒包含第一试剂、第二试剂和复合稳定剂,其中第一试剂组成为:0.1M Tris Buffer pH7.6,鸟嘌呤盐酸盐0.1M,尿酸酶40U/L,过氧化氢酶17KU/L,4-氨基安替比林0.05mM,第二试剂组成为:0.1M Tris Buffer pH6.5,黄嘌呤氧化酶(XOD)240U/L,过氧化物酶(POD)8KU/L,超氧化物歧化酶(SOD)24KU/L,TOOS 0.1M,叠氮钠3.2mM,第一试剂与第二试剂的体积比为4:1。
在GUA测定试剂盒中XOD因为其特异性不高,对多种底物均可发生催化反应,因此其用量需控制在一定的低浓度范围内,这样既可以正常地与黄嘌呤进行氧化反应,又能尽量减少与鸟嘌呤等物质的非特异性反应,降低了测定误差。
所述复合稳定剂中各物质的质量浓度如下:庆大霉素0.002g/L~1.0g/L、牛血清白蛋白(BSA)0.1g/L~10g/L、甘露醇10g/L~100g/L。
优选地,复合稳定剂中各组分的质量浓度如下:庆大霉素0.1g/L~0.5g/L、牛血清白蛋白(BSA)1g/L~8g/L、甘露醇20g/L-80g/L。
优选地,复合稳定剂中各组分的质量浓度如下:庆大霉素0.2g~0.3g/L、牛血清白蛋白(BSA)2g~5g/L、甘露醇30g/L-50g/L。
其中庆大霉素、牛血清白蛋白和甘露醇的含量升高时,其在液态试剂中的溶解性变差,不会再有稳定效果上的提升,还会造成原料浪费,复合稳定剂制造成本升高,含量较低时不能达到稳定液态GUA测试用试剂盒中工具酶的效果,其活性在时间推移下仍会持续降低。
所述庆大霉素具有以下结构:
庆大霉素携带的电荷与XOD酶蛋白表面氨基酸侧链的电荷存在静电作用,通过这种静电作用可以保护XOD酶蛋白的立体结构,防止酶蛋白分子构象发生变换;牛血清白蛋白在溶液中可形成胶体,所述胶体通过共价键连接在XOD酶蛋白表面形成覆盖层,以稳定XOD酶蛋白的立体结构;甘露醇属于非极性溶剂,可通过甘露醇的修饰作用,增加XOD酶蛋白的疏水作用以维持其立体空间结构,通过复合稳定剂中三种物质的共同作用,能防止XOD在保存期间失活,延长鸟嘌呤脱氨酶活性测定试剂盒的稳定期。
实施例1
配置GUA测定用试剂,将配置好的试剂在开盖状态下于8℃储存预定天数,所述试剂为双试剂,具体组成如下:
第一试剂:
第二试剂:
以不添加复合稳定剂的GUA测定用试剂为对照组,分别在对照组中添加不同组分含量的复合稳定剂形成各实验组,分别检测对照组与各实验组的GUA测定用试剂在预定天数测定兔肝提取物中GUA时的活性。
检测使用日立7600-010型全自动生化分析仪完成,试剂仓的储藏温度为2-8℃,检测过程中各参数如下:样本20μL、第一试剂240μL、第二试剂60μL、反应时间10min、主波长570nm、测定方法-速率法,具体检测过程如下:
1、将240μL第一试剂添加至20μL样品中搅拌,然后在37℃下加热5min;
2、加入60μL第二试剂并搅拌,使用日立7600-010型全自动生化分析仪在2min-5min内测量37℃下、570nm波长处的吸光度变化量。
用生理盐溶液做试剂空白,并根据醌染料的微摩尔消光系数(ε=18.44x10-3μM- 1cm-1)计算出的转化系数和吸光度变化量计算鸟嘌呤脱氨酶的活性,计算公式如下:鸟嘌呤脱氨酶的活性GUA(U/L)=F·ΔA,其中Tv为反应体系中液体的总量,即样本、第一试剂、第二试剂的体积总和,Ts为样本的体积,ε为TOOS的摩尔消光系数,ε=18.44x10-3μM-1cm-1,D为比色杯光径,日立分析仪自动转换为1,F为酶活性计算因数,ΔA为吸光度的变化率/min。
检测结果如表2所示:
表2对照组和实验组的活性检测结果
由表2的检测结果可知,不添加复合稳定剂时,GUA活性的测定值随时间的推移呈现显著下降趋势,实验组1和对照组的活性检测几乎没有变化,即复合稳定剂中各组分的含量较低时,其对GUA测定试剂盒没有稳定作用,添加实验组2~实验组6所述的复合稳定剂均能起到稳定GUA测定试剂盒的作用,GUA活性的测量值即使在22天以后也没有大幅度降低,说明使用本发明所述复合稳定剂能够长时间稳定地检测鸟嘌呤脱氨酶的活性,对延长GUA测定试剂盒的稳定期具有一定意义,随着复合稳定剂中各组分浓度的增加,其稳定作用没有更高的变化,且各组分的含量较高时,其在测定试剂中的溶解度降低,即部分物质没有参与反应,造成了浪费。
实施例2
在对照组试剂的基础上,添加实验组3所述的复合稳定剂,将各试剂在2-8℃下避光密封冷藏,在预定天数检测各试剂的中GUA活性值,检测结果如表3和图1所示:
表3鸟嘌呤脱氨酶的活性检测结果
由表3和图1可知在GUA测定用试剂中加入本发明实施例所述的复合稳定剂,可以延长GUA测定试剂盒的稳定期限,使其在12个月内仍能保持良好的活性,方便对其进行存储和推广使用。
实施例3
分别在对照组试剂中添加不同组分含量的复合稳定剂形成各对照组,检测各试剂的稳定活性,检测结果如表4所示:
表4对照组与各实验组的GUA活性检测结果
实验组7添加的复合稳定剂为2.0g/L、甘露醇30.0g/L,实验组8添加的复合稳定剂为庆大霉素0.2g/L,使用实施例1所述仪器、方法对兔肝组织提取物进行检测;由表4可知,分别添加庆大霉素、牛血清白蛋白和甘露醇均能对GUA测定试剂盒的活性进行稳定,但三者联合使用时稳定效果最为显著,这是因为单一稳定剂仅对目标酶具有较好的稳定作用,对其他酶的稳定效果不明显或具有相反的抑制作用,而GUA测定试剂盒中包含多种酶,单独添加庆大霉素、牛血清白蛋白或甘露醇,会使GUA测定试剂盒中的稳定剂彼此抵消,或对一种酶表现出稳定作用,对另一种酶表现出抑制作用,因此本发明实施例根据GUA试剂盒的组合物成分,有选择性的避开相互干扰的稳定剂组分,采用庆大霉素、牛血清白蛋白和甘露醇作为组分,这几种组分之间无干扰作用,能够在稳定目标酶-XOD的同时,对其他工具酶的稳定进行加成补充,更进一步提高GUA试剂盒的稳定性。
本说明书中的各个实施例均采用相关的方式描述,各个实施例之间相同相似的部分互相参见即可,每个实施例重点说明的都是与其他实施例的不同之处。尤其,对于系统实施例而言,由于其基本相似于方法实施例,所以描述的比较简单,相关之处参见方法实施例的部分说明即可。
以上所述仅为本发明的较佳实施例而已,并非用于限定本发明的保护范围。凡在本发明的精神和原则之内所作的任何修改、等同替换、改进等,均包含在本发明的保护范围内。
Claims (5)
1.复合稳定剂,其特征在于,由各组分按以下质量浓度组成:庆大霉素0.002g/L~1.0g/L、牛血清白蛋白0.1g/L~10g/L、甘露醇10g/L~100g/L。
2.根据权利要求1所述的复合稳定剂,其特征在于,由各组分按以下质量浓度组成:庆大霉素0.1g/L~0.5g/L、牛血清白蛋白1g/L~8g/L、甘露醇20g/L~80g/L。
3.根据权利要求1所述的复合稳定剂,其特征在于,由各组分按以下质量浓度组成:庆大霉素0.2g/L~0.3g/L、牛血清白蛋白2g/L~5g/L、甘露醇30g/L~50g/L。
4.含有如权利要求1~3任一项所述复合稳定剂的鸟嘌呤脱氨酶测定试剂盒,其特征在于,鸟嘌呤脱氨酶测定试剂盒包含第一试剂、第二试剂和复合稳定剂,其中第一试剂组成为:0.1M Tris Buffer pH7.6,鸟嘌呤盐酸盐0.1M,尿酸酶40U/L,过氧化氢酶17KU/L,4-氨基安替比林0.05mM;
第二试剂组成为:0.1M Tris Buffer pH6.5,黄嘌呤氧化酶(XOD)240U/L,过氧化物酶(POD)8KU/L,超氧化物歧化酶(SOD)24KU/L,TOOS 0.1M,叠氮钠3.2mM。
5.根据权利要求4所述的鸟嘌呤脱氨酶测定试剂盒,其特征在于,所述第一试剂与第二试剂的体积比为4:1。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0367271A2 (en) * | 1988-11-04 | 1990-05-09 | Sumitomo Seika Chemicals Co., Ltd. | Improved red blood cell preservative solution |
CN1446925A (zh) * | 2003-03-24 | 2003-10-08 | 肖洪武 | 稳定的单一液态酶比色试剂及其应用 |
JP2004242525A (ja) * | 2003-02-12 | 2004-09-02 | Toyobo Co Ltd | 酵素の安定化方法 |
CN101469344A (zh) * | 2007-12-29 | 2009-07-01 | 上海铂锦诊断用品有限公司 | 一种稳定的酶法测定唾液酸液体双试剂及其应用 |
CN104745673A (zh) * | 2015-04-28 | 2015-07-01 | 山东博科生物产业有限公司 | 一种稳定的5’-核糖核苷酸水解酶检测试剂盒 |
CN112481355A (zh) * | 2020-11-16 | 2021-03-12 | 武汉市长立生物技术有限责任公司 | 液体型凝血酶原时间测定试剂盒及其制备方法 |
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Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0367271A2 (en) * | 1988-11-04 | 1990-05-09 | Sumitomo Seika Chemicals Co., Ltd. | Improved red blood cell preservative solution |
JP2004242525A (ja) * | 2003-02-12 | 2004-09-02 | Toyobo Co Ltd | 酵素の安定化方法 |
CN1446925A (zh) * | 2003-03-24 | 2003-10-08 | 肖洪武 | 稳定的单一液态酶比色试剂及其应用 |
CN101469344A (zh) * | 2007-12-29 | 2009-07-01 | 上海铂锦诊断用品有限公司 | 一种稳定的酶法测定唾液酸液体双试剂及其应用 |
CN104745673A (zh) * | 2015-04-28 | 2015-07-01 | 山东博科生物产业有限公司 | 一种稳定的5’-核糖核苷酸水解酶检测试剂盒 |
CN112481355A (zh) * | 2020-11-16 | 2021-03-12 | 武汉市长立生物技术有限责任公司 | 液体型凝血酶原时间测定试剂盒及其制备方法 |
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