CN116482090A - 一种新型尿酸检测试纸 - Google Patents
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Abstract
本发明公开了一种新型尿酸检测试纸,包括以下层次结构:第一层为设有两个直径为1cm圆孔的特质纸,其中一个圆孔内负载有酶片,另一个圆孔为观察孔;第二层由吸水纸和直径为1cm的底物片组成;第三层为设有两个直径为1cm圆孔的特质纸,其中一个圆孔负载有纳米酶片,另一圆孔位置与上层底物片相对应。本发明提供了一种新型尿酸检测试纸,其具有结构简单、制备方便、成本低廉、灵敏度高、特异性强、稳定性好等优点,可用于快速、准确地检测痛风患者的尿酸水平,为痛风的诊断和治疗提供依据;本发明采用了纳米材料作为纳米酶片上的催化剂,其具有类过氧化物酶活性和荧光性质,可同时实现过氧化氢的分解和荧光信号的输出,提高了检测效率和灵敏度。
Description
技术领域
本发明涉及检测试纸技术领域,具体为一种新型尿酸检测试纸。
背景技术
尿酸是人体代谢的终产物之一,它在血液和尿液中的含量与人体健康密切相关。尿酸水平过高或过低都可能导致一些疾病,如痛风、肾结石、高血压、心脏病等。因此,快速、准确、便捷地检测尿酸含量对于疾病的预防和治疗具有重要意义。
目前,检测尿酸含量的方法主要有两类:一类是临床方法,如生化仪法、电极法等,这类方法具有较高的准确性和灵敏度,但是需要专业人员、复杂的仪器设备和标准化的实验室条件,不适合于现场或家庭使用;另一类是非临床方法,如比色法、荧光法等,这类方法具有较低的成本、简单的操作和便携式的特点,但是受到各种干扰因素的影响,其准确性和灵敏度较低。
为了克服现有方法的缺陷,提高尿酸检测的效率和可靠性,本发明提出了一种基于纳米酶催化氧化机理开发一种用于检测痛风患者的试剂条。
发明内容
本发明的目的在于提供一种新型尿酸检测试纸,具备成本低、制备简单、稳定性好的优点,解决了背景技术中的问题。
为实现上述目的,本发明提供如下技术方案:一种新型尿酸检测试纸,包括以下层次结构:
第一层为设有两个直径为1cm圆孔的特质纸,其中一个圆孔内负载有酶片,另一个圆孔为观察孔;
第二层由吸水纸和直径为1cm的底物片组成;
第三层为设有两个直径为1cm圆孔的特质纸,其中一个圆孔负载有纳米酶片,另一圆孔位置与上层底物片相对应;
其中,所述酶片是含有尿酸酶的纸片,所述底物片是含有OPD底物的纸片,所述纳米酶片是具有类过氧化物酶活性和荧光性质的纳米材料。
优选的,所述纳米材料由铁钴氮共掺杂碳点(Fe,Co,N-CDs)组成。
优选的,所述酶片、底物片和纳米酶片均通过戊二醛与壳聚糖载体膜共价交联而固定化。
优选的,所述包括以下步骤:
S1、加入样品溶液:
将样品溶液滴加在酶片(3)上;
S2、生成过氧化氢:
样品溶液与酶片(3)上的尿酸酶发生反应,将尿酸催化为过氧化氢和尿素,反应式如下:
C5H4N4O3+2H2O+O2尿酸酶H2O2+C4H6N4O3+CO2;
S3、催化过氧化氢分解:
折叠试剂条,使底物片和纳米酶片接触,并与纳米酶片上的纳米材料发生反应,催化过氧化氢分解生成羟基自由基,反应式如下:H2O2纳米酶2OH·
S4、生成DAP:
折叠试剂条,使酶片和底物片接触,并与底物片发生反应,生成DAP,反应式如下:OPD+2OH·DAP+2H2O;
S5、观察颜色变化:试剂条颜色由无色变为黄色,在第一层观察孔观察颜色变化。
优选的,所述样品溶液为血清或尿液。
优选的,所述颜色变化与尿酸含量呈线性关系。
与现有技术相比,本发明的有益效果如下:
1、本发明提供了一种新型尿酸检测试纸,其具有结构简单、制备方便、成本低廉、灵敏度高、特异性强、稳定性好等优点,可用于快速、准确地检测痛风患者的尿酸水平,为痛风的诊断和治疗提供依据;
2、本发明采用了纳米材料作为纳米酶片上的催化剂,其具有类过氧化物酶活性和荧光性质,可同时实现过氧化氢的分解和荧光信号的输出,提高了检测效率和灵敏度;
3、本发明通过戊二醛与壳聚糖载体膜共价交联的方式固定化了酶片、底物片和纳米酶片上的生物分子和纳米材料,有效地防止了其在检测过程中的脱落或流失,保证了检测结果的准确性和可重复性;
4、本发明采用了一种简单易行的折叠式试剂条设计,使得检测过程中只需将试剂条按照顺序折叠即可实现各层之间的反应和信号转换,无需额外的仪器或操作,方便了用户的使用和携带;
5、本发明提供了一种使用新型尿酸检测试纸检测痛风患者的方法,其具有操作简单、反应快速、结果直观等优点,可通过观察试剂条颜色变化来判断尿酸含量,并且颜色变化与尿酸含量呈线性关系,具有较高的相关系数,可进行定量分析。
附图说明
图1为本发明的尿酸检测机理;
图2为本发明铁钴氮共掺杂碳点-OPD-H2O2传感器分析尿酸原理;
图3为本发明尿酸检测试纸条具体使用示意图;
图4为本发明一种新型尿酸检测试纸使用方法的流程图;
图5为本发明一种新型尿酸检测试纸的立体结构示意图。
图中:1、特质纸;2、吸水纸;3、酶片;4、底物片;5、纳米酶片。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整的描述,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。
实施例一,本发明提供一种技术方案:一种新型尿酸检测试纸,包括以下层次结构:
第一层为设有两个直径为1cm圆孔的特质纸1,其中一个圆孔内负载有酶片3,另一个圆孔为观察孔;
第二层由吸水纸2和直径为1cm的底物片4组成;
第三层为设有两个直径为1cm圆孔的特质纸1,其中一个圆孔负载有纳米酶片5,另一圆孔位置与上层底物片4相对应;
其中,所述酶片3是含有尿酸酶的纸片,所述底物片4是含有OPD底物的纸片,所述纳米酶片5是具有类过氧化物酶活性和荧光性质的纳米材料。
其中第一层上的观察孔以及第三层上与上层底物片4相对应的圆孔上均附有保护膜,使用时,将其撕开。
实施例一的工作原理:
本发明提供了一种新型尿酸检测试纸,其采用了一种简单易行的折叠式试剂条设计,使得检测过程中只需将试剂条按照顺序折叠即可实现各层之间的反应和信号转换,无需额外的仪器或操作,方便了用户的使用和携带。本发明还通过戊二醛与壳聚糖载体膜共价交联的方式固定化了酶片3、底物片4和纳米酶片5上的生物分子和纳米材料,有效地防止了其在检测过程中的脱落或流失,保证了检测结果的准确性和可重复性;
进一步的,所述纳米材料由铁钴氮共掺杂碳点(Fe,Co,N-CDs)组成;
进一步的,所述酶片3、底物片4和纳米酶片5均通过戊二醛与壳聚糖载体膜共价交联而固定化。
本发明通过戊二醛与壳聚糖载体膜共价交联的方式固定化了酶片、底物片和纳米酶片上的生物分子和纳米材料,有效地防止了其在检测过程中的脱落或流失,保证了检测结果的准确性和可重复性。本发明还利用了壳聚糖载体膜的生物相容性和生物降解性,减少了对环境的污染和对人体的不良影响。
实施例二,一种使用一种新型尿酸检测试纸的方法,包括以下步骤:
S1、加入样品溶液:将样品溶液滴加在酶片3上;
S2、生成过氧化氢:样品溶液与酶片3上的尿酸酶发生反应,将尿酸催化为过氧化氢和尿素,反应式如下:C5H4N4O3+2H2O+O2尿酸酶H2O2+C4H6N4O3+CO2;
S3、催化过氧化氢分解:折叠试剂条,使底物片4和纳米酶片5接触,并与纳米酶片5上的纳米材料发生反应,催化过氧化氢分解生成羟基自由基,反应式如下:H2O2纳米酶2OH·;
S4、生成DAP:折叠试剂条,使酶片3和底物片4接触,并与底物片4发生反应,生成DAP,反应式如下:OPD+2OH·DAP+2H2O;
S5、观察颜色变化:试剂条颜色由无色变为黄色,在第一层观察孔观察颜色变化;
DAP具有在I580/I475处存在线性关系的荧光特性;
本发明提供了一种使用新型尿酸检测试纸检测痛风患者的方法,其具有操作简单、反应快速的效果;
使用新型尿酸检测试纸的具体方法步骤,包括五个步骤,分别是加入样品溶液、生成过氧化氢、催化过氧化氢分解、生成DAP、观察颜色变化。
利用尿酸酶和纳米材料的催化作用,实现尿酸的快速检测,通过颜色变化或荧光信号的强度判断尿酸含量的高低。
反应过程中,尿酸被催化氧化生成H2O2,H2O2催化氧化OPD生成DAP,并且产生的DAP可见的颜色改变(从无色到黄色)在360nm的激发峰下在580nm处发射出黄色荧光,同时猝灭该碳点在475nm的蓝色荧光发射峰,导致发射强度比(I580/I475)与尿酸浓度存在着线性关系。
OPD可通过内滤效应(IFE)有效地猝灭纳米材料的荧光,从而产生新的发射峰,导致发射强度比(IOPD/IN-CDs)变化;
关于内滤效应(IFE),内滤效应是一种荧光分析中常见的现象,它指的是当溶液中存在能够吸收荧光物质的激发光或发射光(或两者都吸收)的物质时,会导致荧光物质的荧光强度降低。内滤效应可以分为初级内滤效应和次级内滤效应,前者是指荧光物质自身或其他物质对激发光的吸收,后者是指荧光物质自身或其他物质对发射光的吸收;
将纳米材料、过氧化物酶底物和尿酸酶分别固定在不同的纸片上,形成可折叠的三层结构,方便携带和使用,同时覆盖有透明盖膜,保护其稳定性、活性和防潮。
进一步的,所述样品溶液为血清或尿液。
本发明可使用血清或尿液作为样品溶液,无需进行复杂的预处理或稀释,可直接加入试剂条进行检测,提高了检测的便捷性和实用性。
利用纳米材料的催化作用和过氧化物酶底物的氧化反应,实现了尿酸的快速检测,避免了传统方法中需要的复杂仪器、耗时操作和高成本。
进一步的,纳米材料选自铁钴氮共掺杂碳点(Fe,Co,N-CDs);
本发明通过戊二醛与壳聚糖载体膜共价交联的方式固定化了酶片、底物片和纳米酶片上的生物分子和纳米材料,有效地防止了其在检测过程中的脱落或流失,保证了检测结果的准确性和可重复性。本发明还利用了壳聚糖载体膜的生物相容性和生物降解性,减少了对环境的污染和对人体的不良影响;
进一步的,所述颜色变化与尿酸含量呈线性关系;
本发明实现了颜色变化与尿酸含量之间的线性关系,且具有较高的相关系数,可进行定量分析。
这一部分内容描述了尿酸检测试纸的检测原理,即颜色变化与尿酸含量呈线性关系,也就是说,尿酸含量越高,颜色变化越明显,反之亦然。
这一部分内容的目的是利用颜色变化作为检测信号,实现尿酸含量的定量分析,通过比较颜色变化或荧光信号的强度与标准曲线,计算出尿酸含量的数值。
利用纳米材料的催化作用和过氧化物酶底物的氧化反应,实现了尿酸的快速检测,避免了传统方法中需要的复杂仪器、耗时操作和高成本。
描述了尿酸检测试纸的检测原理,即颜色变化与尿酸含量呈线性关系,也就是说,尿酸含量越高,颜色变化越明显,反之亦然;
利用颜色变化作为检测信号,实现尿酸含量的定量分析,通过比较颜色变化或荧光信号的强度与标准曲线,计算出尿酸含量的数值。
采用比色/荧光双信号传感器,既可以利用肉眼观察颜色变化,也可以利用仪器测量荧光信号,提高了检测的灵敏度和准确度;
尽管已经示出和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。
Claims (6)
1.一种新型尿酸检测试纸,其特征在于:包括以下层次结构:
第一层为设有两个直径为1cm圆孔的特质纸(1),其中一个圆孔内负载有酶片(3),另一个圆孔为观察孔;
第二层由吸水纸(2)和直径为1cm的底物片(4)组成;
第三层为设有两个直径为1cm圆孔的特质纸(1),其中一个圆孔负载有纳米酶片(5),另一圆孔位置与上层底物片(4)相对应;
其中,所述酶片(3)是含有尿酸酶的纸片,所述底物片(4)是含有OPD底物的纸片,所述纳米酶片(5)是具有类过氧化物酶活性和荧光性质的纳米材料。
2.根据权利要求1所述的一种新型尿酸检测试纸,其特征在于:所述纳米材料由铁钴氮共掺杂碳点组成。
3.根据权利要求2所述的一种新型尿酸检测试纸,其特征在于:所述酶片(3)、底物片(4)和纳米酶片(5)均通过戊二醛与壳聚糖载体膜共价交联而固定化。
4.一种使用权利要求3所述的一种新型尿酸检测试纸的方法,其特征在于:包括以下步骤:
S1、加入样品溶液:
将样品溶液滴加在酶片(3)上;
S2、生成过氧化氢:
样品溶液与酶片(3)上的尿酸酶发生反应,将尿酸催化为过氧化氢和尿素,反应式如下:C5H4N4O3+2H2O+O2尿酸酶H2O2+C4H6N4O3+CO2;
S3、催化过氧化氢分解:
折叠试剂条,使底物片(4)和纳米酶片(5)接触,并与纳米酶片(5)上的纳米材料发生反应,催化过氧化氢分解生成羟基自由基,反应式如下:H2O2纳米酶2OH·;
S4、生成DAP:
折叠试剂条,使酶片(3)和底物片(4)接触,并与底物片(4)发生反应,生成DAP,反应式如下:OPD+2OH·DAP+2H2O;
S5、观察颜色变化:
试剂条颜色由无色变为黄色,在第一层观察孔观察颜色变化。
5.根据权利要求4所述的方法,其特征在于:所述样品溶液为血清或尿液。
6.根据权利要求5所述的方法,其特征在于:所述颜色变化与尿酸含量呈线性关系。
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