CN114235790A - 体液中乳酸含量检测试纸的制备方法 - Google Patents
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Abstract
本发明涉及一种体液中乳酸含量检测试纸的制备方法,包括以下步骤:配制GOODS缓冲液:将溶剂加入纯化水中用氢氧化钠溶液调PH至6.0‑7.0;配制乳酸试纸处理液:量取GOODS缓冲液按顺序加入稳定剂、CHAPS、TOOS、乳酸氧化酶、过氧化物酶,搅拌至完全溶解得乳酸试纸处理液;用滤纸或棉纤维做为试剂载体,浸乳酸试纸处理液,烘干,然后裁切成一定尺寸的小块,粘贴在塑料基片上,放入密封的塑料瓶或者铝箔袋中即得。本发明利用乳酸氧化酶和过氧化物酶,通过一系列反应显色测试乳酸的含量,配方中使用的新型的保护剂,使用底物CHAPS和TOOS在该保护剂的作用下,该两种底物稳定存在于溶液中,实现了配制和制作一步法,不需要酶溶液和底物溶液分两次配制及浸渍烘干。
Description
技术领域
本发明属于体外诊断试纸技术领域,具体涉及体液中乳酸含量检测试纸的制备方法。
背景技术
下生殖道感染属于妇科常见病,易导致流产、***不育、***甚至***,现已成为发展中国家面临的重要公共卫生问题。
***炎以各种原因导致的***菌群失调及以***微生态平衡破坏为首要影响因素。近年来临床病例显示常见妇***道炎多数是由大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌、白色念珠菌、伤寒杆菌和肠球菌引起的。乳酸杆菌是健康妇***道中的正常菌种和优势菌群,它是一种微需氧菌群,与***菌群之间形成微生态平衡并制约其他菌的生长。乳酸杆菌的代谢产物(乳酸)可以降低***pH值,维持***的生态平衡,作为妇***道的清洗剂,无毒、无副作用,在保持***清洁等方面起重要作用。研究表明各种***炎患者的乳酸杆菌数量明显下降,乳酸杆菌的替代、排斥和竞争等保护作用减少或消失,寄生在***的支原体、真菌、加德纳杆菌、或其他外来病原微生物在***中大量繁殖,引起各种***炎的发作,***中乳酸杆菌数量的减少在各种***炎的发病过程中起着非常关键的作用。
专利号为CN201811207828.9的发明专利提供了一种乳酸检测干化学试纸及其制备方法,属于体外诊断试纸技术领域。解决了如何提供一种简单、快捷、安全、有效,对***炎的诊断具有很高的敏感度和特异性的乳酸检测干化学试纸及其制备方法的问题。本发明的乳酸检测干化学试纸,为分散有底物、酶、螯合剂、保护剂和缓冲物的滤纸;其中,底物为联苯胺类或联苯胺类衍生物;酶为乳酸氧化酶和过氧化物酶。该干化学试纸可一步测定乳酸含量,简单、快捷、安全、有效、敏感度高、特异性好。
但是上述专利中的产品制备时采用A、B两遍液制作,过程繁琐,人工成本增高而且多一遍制作过程,会导致产品出现误差的几率加大。
发明内容
为解决现有技术中存在的问题,本发明提供一种体液中乳酸含量检测试纸的制备方法,按照先后顺序包括以下步骤:
(1)配制GOODS缓冲液:将溶剂加入纯化水中用氢氧化钠溶液调PH至6.0-7.0;
(2)配制乳酸试纸处理液:量取GOODS缓冲液按顺序加入稳定剂、CHAPS、TOOS、乳酸氧化酶、过氧化物酶,搅拌至完全溶解得乳酸试纸处理液;
(3)用滤纸或棉纤维做为试剂载体,浸乳酸试纸处理液,烘干,然后裁切成一定尺寸的小块,粘贴在塑料基片上,放入密封的塑料瓶或者铝箔袋中即得。
优选的是,步骤(1)中的溶剂为2-(N-吗啉代)乙磺酸、2[(2-氨基-2氧代乙基)氨基]乙磺酸、哌嚓-N,N-二(2-乙磺酸)或3-(N-吗啉代)-2-羟基丙磺酸中的一种。
在上述任一方案中优选的是,所述稳定剂为:牛血清白蛋白和聚乙二醇8000的混合溶液。
在上述任一方案中优选的是,GOODS缓冲液浓度在0.1M-1M之间,PH范围为6.0-7.0。
在上述任一方案中优选的是,牛血清白蛋白的浓度0.1%-1%,聚乙二醇8000的浓度0.05%-0.5%。
在上述任一方案中优选的是,步骤(3)中,烘干的条件为:用烘箱60℃-80℃烘干10分钟-20分钟。
在上述任一方案中优选的是,选用的保护剂是二硫代苏糖醇(DDT),乳酸氧化酶的末端硫原子在溶液中趋向于形成二聚体,而这种二聚体很大程度的降低了偶联反应的效率,在乳酸氧化酶溶液中同时加入DDT可以有效降低二聚化,从而提高了乳酸氧化酶的反应效率。
本发明利用乳酸氧化酶和过氧化物酶,通过一系列反应显色测试乳酸的含量,配方中使用的新型的保护剂,使用底物CHAPS和TOOS在该保护剂的作用下,该两种底物稳定存在于溶液中,实现了配制和制作一步法,不需要酶溶液和底物溶液分两次配制及浸渍烘干。
具体实施方式
为了更进一步了解本发明的发明内容,下面将结合具体实施例详细阐述本发明。
实施例一
1.配制GOODS缓冲液;取2-(N-吗啉代)乙磺酸、2[(2-氨基-2氧代乙基)氨基]乙磺酸、哌嚓-N,N-二(2-乙磺酸)、3-(N-吗啉代)-2-羟基丙磺酸等其中一种试剂作为溶质加入纯化水中用相应浓度氢氧化钠溶液调PH至6.0-7.0;
2.配制乳酸试纸处理液:准确量取GOODS缓冲液按顺序加入牛血清白蛋白、聚乙二醇8000、CHAPS、TOOS、乳酸氧化酶、过氧化物酶等,搅拌完全溶解;
3.浸干操作:将GE公司3mm滤纸用上述溶液浸泡10秒-30秒后,用烘箱60℃-80℃烘干10分钟-20分钟,试纸烘干以后取出;
4.产品组装:将烘干后的试纸裁切成一定尺寸的小块,粘贴在塑料基片上。放入密封的塑料瓶或者铝箔袋中,用13X型分子筛做为干燥剂,室温可存放有效期18个月。
实施例二
1.配制GOODS缓冲液;取2-(N-吗啉代)乙磺酸、2[(2-氨基-2氧代乙基)氨基]乙磺酸、哌嚓-N,N-二(2-乙磺酸)、3-(N-吗啉代)-2-羟基丙磺酸等其中一种试剂作为溶质加入纯化水中用相应浓度氢氧化钠溶液调PH至6.0-7.0;
2.配制乳酸试纸处理液:准确量取GOODS缓冲液按顺序加入牛血清白蛋白、聚乙二醇8000、CHAPS、TOOS、乳酸氧化酶、过氧化物酶等,搅拌完全溶解;
3.浸干操作:将GE公司3mm滤纸用上述溶液浸泡10秒-30秒后,用烘箱60℃-80℃烘干10分钟-20分钟,试纸烘干以后取出;
4.产品组装:将烘干后的试纸裁切成一定尺寸的小块,粘贴在塑料基片上。放入密封的塑料瓶或者铝箔袋中,用13X型分子筛做为干燥剂,室温可存放有效期18个月。
实施例三
1.配制GOODS缓冲液;取2-(N-吗啉代)乙磺酸、2[(2-氨基-2氧代乙基)氨基]乙磺酸、哌嚓-N,N-二(2-乙磺酸)、3-(N-吗啉代)-2-羟基丙磺酸等其中一种试剂作为溶质加入纯化水中用相应浓度氢氧化钠溶液调PH至6.0-7.0;
2.配制乳酸试纸处理液:准确量取GOODS缓冲液按顺序加入牛血清白蛋白、聚乙二醇8000、CHAPS、TOOS、乳酸氧化酶、过氧化物酶等,搅拌完全溶解;
3.浸干操作:将GE公司3mm滤纸用上述溶液浸泡10秒-30秒后,用烘箱60℃-80℃烘干10分钟-20分钟,试纸烘干以后取出;
4.产品组装:将烘干后的试纸裁切成一定尺寸的小块,粘贴在塑料基片上。放入密封的塑料瓶或者铝箔袋中,用13X型分子筛做为干燥剂,室温可存放有效期18个月。
1.临界值:尿半乳糖浓度1mmol/L,尿半乳糖浓度<1mmol/L阳性,尿半乳糖浓度≥1mmol/L为阴性。
2.阳性判断值的建立:本产品阳性判断值是经180例正常人群验证。
本领域技术人员不难理解,本发明的体液中乳酸含量检测试纸的制备方法包括上述本发明说明书的发明内容和具体实施方式部分所示出的各部分的任意组合,限于篇幅并为使说明书简明而没有将这些组合构成的各方案一一描述。凡在本发明的精神和原则之内,所做的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (6)
1.一种体液中乳酸含量检测试纸的制备方法,按照先后顺序包括以下步骤:
(1)配制GOODS缓冲液:将溶剂加入纯化水中用氢氧化钠溶液调PH至6.0-7.0;
(2)配制乳酸试纸处理液:量取GOODS缓冲液按顺序加入稳定剂、CHAPS、TOOS、乳酸氧化酶、过氧化物酶,搅拌至完全溶解得乳酸试纸处理液;
(3)用滤纸或棉纤维做为试剂载体,浸乳酸试纸处理液,烘干,然后裁切成一定尺寸的小块,粘贴在塑料基片上,放入密封的塑料瓶或者铝箔袋中即得。
2.根据权利要求1所述的体液中乳酸含量检测试纸的制备方法,其特征在于,步骤(1)中的溶剂为2-(N-吗啉代)乙磺酸、2[(2-氨基-2氧代乙基)氨基]乙磺酸、哌嚓-N,N-二(2-乙磺酸)或3-(N-吗啉代)-2-羟基丙磺酸中的一种。
3.根据权利要求1所述的体液中乳酸含量检测试纸的制备方法,其特征在于,所述稳定剂为:牛血清白蛋白和聚乙二醇8000的混合溶液。
4.根据权利要求1所述的体液中乳酸含量检测试纸的制备方法,其特征在于,GOODS缓冲液浓度在0.1M-1M之间,PH范围为6.0-7.0。
5.根据权利要求3所述的体液中乳酸含量检测试纸的制备方法,其特征在于,牛血清白蛋白的浓度0.1%-1%,聚乙二醇8000的浓度0.05%-0.5%。
6.根据权利要求1所述的体液中乳酸含量检测试纸的制备方法,其特征在于,步骤(3)中,烘干的条件为:用烘箱60℃-80℃烘干10分钟-20分钟。
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