CN107884399A - 用于检测红细胞的嘌呤类化合物修饰纳米金材料,制备方法和试剂盒 - Google Patents
用于检测红细胞的嘌呤类化合物修饰纳米金材料,制备方法和试剂盒 Download PDFInfo
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Abstract
本发明涉及一种用于检测红细胞的嘌呤类化合物修饰纳米金材料,制备方法和试剂盒。该材料以纳米金为主体,通过Au‑N键将嘌呤类化合物修饰于纳米金表面,其能够催化过氧化氢使指示剂3,3’,5,5’‑四甲基联苯胺(TMB)变色。Fe2+会对纳米金体系的催化活性产生显著影响,通过TMB的紫外‑可见吸收光谱变化能够定量分析溶液中的Fe2+。Fe2+普遍存在于红细胞中,与卟啉环配位成为血红素的活性中心。因此,应用嘌呤衍生物修饰的纳米金体系可以实现对红细胞的分析,并可用于定量检测尿液中的红细胞,进而发展为一种尿潜血检测试剂盒。本发明设计的尿潜血检测试剂盒具有灵敏度较高,操作简便,反应迅速,稳定性好的优势。
Description
技术领域
本发明属于生物技术领域,具体涉及一种用于检测红细胞的嘌呤类化合物修饰纳米金材料,其制备方法,和基于该材料用于尿潜血检测的试剂盒。
背景技术
在日常生活中,尿潜血是通过尿常规检查出来的,尿常规中有一项红细胞。如果红细胞增多,以其数量来讲,少数就可以称为尿潜血。尿潜血是IGA肾病、肾炎的常见症状,也可能是肾囊肿或多囊肿破裂所为。尿潜血的原因一般归因于以下三项原因,一是炎症,一是结石,再则是肿瘤。关于炎症方面,如肾小球炎、膀胱炎等,除了可能会有血尿发生,尿液检查也会有潜血发生,结石本身不论是肾脏、输尿管或膀胱结石,都可能造成潜血,其他情境如生理期时,便秘也可能造成潜血。另外,肿瘤也会引起潜血,如肾、膀胱、输尿管的良性或恶性肿瘤。因此,检测尿潜血对潜在疾病的发现和及时治疗有重要的意义。
纳米金是一类能够模拟天然过氧化氢酶(HRP)的无机纳米材料,天然的HRP不易提取、存储,同时在强酸碱和高温条件下极易失活。而纳米级别的金溶胶能够克服这些缺陷,可在严苛的条件下依然保持较高的活性,催化过氧化氢使有机指示剂3,3‘,5,5’-四甲基联苯胺(TMB)变色。红细胞可加快反应,产生更深的颜色变化。同时,体系的吸光度同红细胞浓度呈现良好的线性相关。因此,该发明可用于间接检测尿液中红细胞浓度。
发明内容
本发明的目的是一种用于检测红细胞的嘌呤类化合物修饰纳米金材料,其制备方法,和基于该材料用于尿潜血检测的试剂盒。
一种用于检测红细胞的嘌呤类化合物修饰纳米金材料,以纳米金为主体,通过Au-N键将嘌呤类化合物修饰于纳米金表面。
优选的,所述嘌呤类化合物是2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤或6-肼基嘌呤中的任意一种。
一种用于检测红细胞的嘌呤类化合物修饰纳米金材料的制备方法,在冰水浴条件下,按顺序向嘌呤类化合物水溶液中加入吐温80、氯金酸和硼氢化钠进行反应。
优选的,所述嘌呤类化合物同氯金酸摩尔比例在2:1-50:1,硼氢化钠用量同氯金酸质量比在1:10-5:1,吐温80加入量同总溶液的体积比在1:100-1:10。
优选的,所述嘌呤类化合物是2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤或6-肼基嘌呤中的任意一种。
一种试剂盒,使用上述用于检测红细胞的嘌呤类化合物修饰纳米金材料,用于检测尿潜血。
优选的,所述试剂盒中包括:第一玻璃管,含有上述用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液的混合液;第二玻璃管,含有H2O2和TMB溶液的混合液;加入尿液样品后,折断玻璃管进行检测反应。HAc-NaAc缓冲液的pH值范围为2.0-3.2,用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液的体积比为1:1.5-1:4.5,H2O2和TMB溶液的体积比为0.75-1.5:1,H2O2浓度范围为0.1-1mol/L,TMB溶液浓度范围为10-50mM。
一种检测Fe2+的方法,取上述用于检测红细胞的嘌呤类化合物修饰纳米金材料,加入pH值范围为2.0-3.2的HAc-NaAc缓冲液、H2O2、TMB溶液和不同浓度的Fe2+溶液反应,观察颜色变化,并测量紫外吸收。用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液的体积比为1:1.5-1:4.5,H2O2和TMB溶液的体积比为0.75-1.5:1,H2O2浓度范围为0.1-1mol/L,TMB溶液浓度范围为10-50mM。用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液体积之和与H2O2和TMB溶液的体积之和相同。
一种红细胞的检测方法,取上述用于检测红细胞的嘌呤类化合物修饰纳米金材料,加入pH值范围为2.0-3.2的HAc-NaAc缓冲液、H2O2、TMB和不同浓度的红细胞溶液,观察颜色变化,并测量其紫外吸收。用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液的体积比为1:1.5-1:4.5,H2O2和TMB溶液的体积比为0.75-1.5:1,H2O2浓度范围为0.1-1mol/L,TMB溶液浓度范围为10-50mM。用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液体积之和与H2O2和TMB溶液的体积之和相同。
本发明通过合成嘌呤类衍生物修饰的纳米金,利用该物质的高比表面积、高催化活性,通过红细胞的加入能够使其产生更高的过氧化氢催化活性的原理,间接检验尿液中红细胞的存在,同时可根据颜色变化深浅来判断尿潜血的程度。本发明所提供的嘌呤类衍生物修饰的纳米金具有很好的稳定性、催化活性,对红细胞灵敏度高、检测限低的优势,可进一步应用于尿潜血试剂盒。
附图说明
图1是本发明实施例中2,6-二氨基嘌呤纳米金的XPS图。
图2是本发明实施例中2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤、6-肼基嘌呤修饰的纳米金的过氧化氢催化时间变化曲线图。
图3是本发明实施例中2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤、6-肼基嘌呤修饰的纳米金的zeta和DLS图。
图4是本发明实施例中2,6-二氨基嘌呤修饰的纳米金的稳定性可视化图。
图5是本发明实施例中不同浓度的Fe2+在水溶液中催化时间曲线。
图6是本发明实施例中不同浓度的Fe2+在水溶液中对体系催化活性的影响曲线。
图7是本发明实施例中不同浓度的Fe2+在水溶液中,吸光度随浓度线性变化曲线。
图8是本发明实施例中不同浓度的红细胞在水溶液中催化时间曲线。
图9是本发明实施例中不同浓度的红细胞在水溶液中对体系催化活性的影响曲线。
图10是本发明实施例中不同浓度的红细胞在水溶液中,吸光度随浓度线性变化曲线。
图11是本发明实施例中不同浓度的红细胞在尿液中催化时间曲线。
图12是本发明实施例中不同浓度的红细胞在尿液中对体系催化活性的影响曲线。
图13是本发明实施例中不同浓度的红细胞在尿液中,吸光度随浓度线性变化曲线。
图14是本发明实施例中实施例提供的尿潜血试剂盒检测尿液中红细胞步骤可视化图片。
具体实施方式
纳米金,是一种尺寸约3nm大小的金颗粒,具有高电子密度、介电特性和催化作用,研究发现纳米金材料具有过氧化物模拟酶性质,使得其可以像一些天然的过氧化物酶(比如辣根过氧化物酶)一样催化一系列过氧化氢底物显色。本发明是用嘌呤类衍生物修饰的纳米金颗粒具有过氧化氢酶活性,使有机指示剂3,3’,5,5’-四甲基联苯胺(TMB)变色,且对红细胞具有更高的灵敏度和更低的检出限。Fe2+普遍存在于红细胞中,与卟啉环配位成为血红素的活性中心。相比于其他种类阳离子,Fe2+会大大增加体系的催化活性,产生更高的紫外吸收,实验发现红细胞同样能够增加体系的催化活性,且浓度越高,体系的吸光度越高。发明提供的嘌呤类衍生物修饰的纳米金颗粒可间接用于检测尿液中的红细胞。
嘌呤类化合物修饰纳米金材料的制备实施例
嘌呤类化合物配体修饰纳米金的制备:
分别称取22.5mg(0.15mM)的2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤、6-肼基嘌呤,溶于50ml去离子水中,转移至合成瓶。在冰水浴中搅拌5min,加入0.2ml吐温80,继续搅拌5min。随后加入155μL(40g/L)氯金酸(0.015mM),混合均匀后,加入0.3ml10mg/L的硼氢化钠还原剂,溶液变为棕黄色,继续反应45min得到分别由2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤、6-肼基嘌呤修饰的纳米金溶胶。
不同配体修饰纳米金的过氧化氢催化时间变化实验
具体步骤如下:
(1)取1500μLpH值3.0的HAc-NaAc缓冲液、500μL 10mM H2O2、500μL 10mM TMB和500μL制备实施例得到的不同配体修饰的纳米金混合;
(2)分别在反应2min、5min、8min、12min、16min、20min、25min、30min、45min、60min和90min后测溶液在655nm处的紫外吸收;如图2中所示。
不同配体修饰纳米金的稳定性可视化图
具体步骤如下:
分别取2.5ml制备实施例得到的五种嘌呤类化合物修饰纳米金于样品小瓶中在0h、2h、5h、10h、1d、2d、3d进行连续的拍照观察至肉眼发现纳米金团聚。例如,图4是2,6-二氨基嘌呤修饰的纳米金的稳定性可视化图。
不同浓度的红细胞在缓冲溶液中催化时间曲线
具体步骤如下:
(1)取数只5ml小管,分别加入500μL制备实施例得到的2,6-二氨基嘌呤修饰的纳米金、1500μLpH值3.0的HAc-NaAc缓冲液、500μL 10mM H2O2、500μL 5mM TMB和500μL含不同浓度的红细胞的水溶液;
(2)分别在反应2min、5min、8min、12min、16min、20min、25min、30min、45min、60min和90min后测溶液在655nm处的紫外吸收;如图8中所示。
不同浓度的红细胞在缓冲溶液中对催化活性影响实验
具体步骤如下:
(1)取数只5ml小管,分别加入500μL制备实施例得到的2,6-二氨基嘌呤修饰的纳米金、1500μLpH值3.0的HAc-NaAc缓冲液、500μL 10mM H2O2、500μL 2.5mM TMB和500μL含不同浓度的红细胞的水溶液;
(2)等待30min观察颜色变化,并测在655nm处紫外吸收;如图9中所示。
不同浓度的红细胞在尿液中催化时间曲线
具体步骤如下:
(1)取数只5ml小管,分别加入500μL制备实施例得到的2,6-二氨基嘌呤修饰的纳米金、1500μLpH值3.0的HAc-NaAc缓冲液、500μL 10mM H2O2、500μL 5mM TMB和500μL含不同浓度的红细胞的尿液;
(2)分别在反应2min、5min、8min、12min、16min、20min、25min、30min、45min、60min和90min后测溶液在655nm处的紫外吸收;如图11中所示。
不同浓度的红细胞在尿液中对催化活性影响实验
具体步骤如下:
(1)取数只5ml小管,分别加入500μL制备实施例得到的2,6-二氨基嘌呤修饰的纳米金、1500μLpH值3.0的HAc-NaAc缓冲液、500μL 10mM H2O2、500μL 2.5mM TMB和500μL含不同浓度的红细胞的水溶液;
(2)等待30min观察颜色变化,并测在655nm处紫外吸收;如图12中所示。
尿潜血试剂盒
具体步骤如下:
(1)取一只透明软管中封存有两根玻璃管,第一玻璃管,含有制备实施例得到的2,6-二氨基嘌呤修饰的纳米金和HAc-NaAc缓冲液的混合液;第二玻璃管,含有H2O2和TMB溶液的混合液;2,6-二氨基嘌呤修饰的纳米金和HAc-NaAc缓冲液的体积比1:3,H2O2和TMB溶液的体积比为1:1,双氧水浓度为0.5M,TMB溶液浓度为20mM;两根玻璃管中混合液的体积相同;
(2)加入2ml尿液样品后,弯曲软管以折断内含的玻璃管;
(3)2min后观察溶液颜色变化。例如图14中步骤所示。
可以理解的是,以上是为了阐述本发明的原理和可实施性的示例,本发明并不局限于此。对于本领域内的普通技术人员而言,在不脱离本发明的精神和实质的情况下,可以做出各种变型和改进,这些变型和改进也视为本发明的保护范围。
Claims (9)
1.一种用于检测红细胞的嘌呤类化合物修饰纳米金材料,其特征在于,以纳米金为主体,通过Au-N键将嘌呤类化合物修饰于纳米金表面。
2.根据权利要求1所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料,其特征在于,所述嘌呤类化合物是2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤或6-肼基嘌呤中的任意一种。
3.一种用于检测红细胞的嘌呤类化合物修饰纳米金材料的制备方法,其特征在于,在冰水浴条件下,按顺序向嘌呤类化合物水溶液中加入吐温80、氯金酸和硼氢化钠进行反应。
4.根据权利要求3所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料的制备方法,其特征在于,所述嘌呤类化合物同氯金酸摩尔比例在2:1-50:1,硼氢化钠用量同氯金酸质量比在1:10-5:1,吐温80加入量同总溶液的体积比在1:100-1:10。
5.根据权利要求4所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料的制备方法,其特征在于,所述嘌呤类化合物是2,6-二氨基嘌呤、6-巯基嘌呤、腺嘌呤、嘌呤或6-肼基嘌呤中的任意一种。
6.一种Fe2+的检测方法,其特征在于,该方法具体操作步骤为;取权利要求2所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料,加入pH值范围为2.0-3.2的HAc-NaAc缓冲液、H2O2、TMB溶液和不同浓度的Fe2+溶液反应,观察颜色变化,并测量其紫外吸收。
7.一种红细胞的检测方法,其特征在于,该方法具体操作步骤为;取权利要求2所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料,加入pH值范围为2.0-3.2的HAc-NaAc缓冲液、H2O2、TMB溶液和不同浓度的红细胞溶液,观察颜色变化,并测量其紫外吸收。
8.一种试剂盒,其特征在于,使用权利要求2中所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料,用于检测尿潜血。
9.根据权利要求8所述的试剂盒,所述试剂盒中包括:第一玻璃管,含有权利要求2中所述的用于检测红细胞的嘌呤类化合物修饰纳米金材料和HAc-NaAc缓冲液的混合液;第二玻璃管,含有H2O2和TMB溶液的混合液;加入尿液样品后,折断玻璃管进行检测反应;HAc-NaAc缓冲液的pH值范围为2.0-3.2。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110567953A (zh) * | 2019-10-12 | 2019-12-13 | 山西师范大学 | 用于检测环境水样和血清中Fe2+含量的可视化检测试剂盒及其检测方法 |
CN111537461A (zh) * | 2020-05-28 | 2020-08-14 | 武汉科技大学 | 一种硼簇纳米金检测溶液中腺嘌呤和鸟嘌呤的方法 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004086053A1 (en) * | 2003-03-25 | 2004-10-07 | Stichting Sanquin Bloedvoorziening | Method for the detection of a pathogenic form of a prion protein |
CN201642498U (zh) * | 2010-03-30 | 2010-11-24 | 林庆娟 | 一种自动检测隐形血尿的储尿袋 |
CN105675598A (zh) * | 2016-01-20 | 2016-06-15 | 曲阜师范大学 | 一种基于血红素和纳米金簇的蛋白模拟酶制备方法及应用 |
CN107091926A (zh) * | 2017-03-13 | 2017-08-25 | 广东省生态环境技术研究所 | 一种四环素的检测方法及检测试剂盒 |
CN107976435A (zh) * | 2017-10-27 | 2018-05-01 | 中国农业大学 | 一种基于功能核酸的传感器及其在钠离子检测中的应用 |
-
2017
- 2017-11-08 CN CN201711093692.9A patent/CN107884399B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004086053A1 (en) * | 2003-03-25 | 2004-10-07 | Stichting Sanquin Bloedvoorziening | Method for the detection of a pathogenic form of a prion protein |
CN201642498U (zh) * | 2010-03-30 | 2010-11-24 | 林庆娟 | 一种自动检测隐形血尿的储尿袋 |
CN105675598A (zh) * | 2016-01-20 | 2016-06-15 | 曲阜师范大学 | 一种基于血红素和纳米金簇的蛋白模拟酶制备方法及应用 |
CN107091926A (zh) * | 2017-03-13 | 2017-08-25 | 广东省生态环境技术研究所 | 一种四环素的检测方法及检测试剂盒 |
CN107976435A (zh) * | 2017-10-27 | 2018-05-01 | 中国农业大学 | 一种基于功能核酸的传感器及其在钠离子检测中的应用 |
Non-Patent Citations (2)
Title |
---|
MUSTAFA SALIH HIZIR ET AL: "Multiplexed Activity of perAuxidase: DNA-Capped AuNPs Act as Adjustable Peroxidase", 《ANAL. CHEM.》 * |
NAK HAN JANG: "The Coordination Chemistry of DNA Nucleosides on Gold Nanoparticles as a Probe by SERS", 《BULL. KOREAN CHEM. SOC.》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110567953A (zh) * | 2019-10-12 | 2019-12-13 | 山西师范大学 | 用于检测环境水样和血清中Fe2+含量的可视化检测试剂盒及其检测方法 |
CN110567953B (zh) * | 2019-10-12 | 2022-07-01 | 山西师范大学 | 用于检测环境水样和血清中Fe2+含量的可视化检测试剂盒及其检测方法 |
CN111537461A (zh) * | 2020-05-28 | 2020-08-14 | 武汉科技大学 | 一种硼簇纳米金检测溶液中腺嘌呤和鸟嘌呤的方法 |
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