CN111441022A - 一种sers增强的新冠病毒检测芯片的制备方法及其产品和应用 - Google Patents

一种sers增强的新冠病毒检测芯片的制备方法及其产品和应用 Download PDF

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CN111441022A
CN111441022A CN202010313913.4A CN202010313913A CN111441022A CN 111441022 A CN111441022 A CN 111441022A CN 202010313913 A CN202010313913 A CN 202010313913A CN 111441022 A CN111441022 A CN 111441022A
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nano
sers
detection
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oscillating
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崔大祥
朱君
徐艳
杨迪诚
金彩虹
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Shanghai National Engineering Research Center for Nanotechnology Co Ltd
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Abstract

本发明提供一种SERS增强的新冠病毒检测芯片的制备方法及其产品和应用，通过制备纳米银/纳米氧化锌/纳米金的SERS基底，从而增加荧光分子的发光强度。可以应用于新冠病毒的检测中，采用WHO公布的引物和探针，在SERS基底连接荧光探针，实现高灵敏检测，提高检测效率和准确度的同时降低了成本，可用于新型冠状病毒的快速应急检测。本发明制备的芯片大大增强了小分子的拉曼信号，光学强度高，检测灵敏度高，检测限低。本发明制备过程简单、反应易控制，稳定性好、可产业化，在新冠病毒的检测领域具有广泛的应用前景。

Description

一种SERS增强的新冠病毒检测芯片的制备方法及其产品和 应用
技术领域
本发明属于纳米材料的功能化及芯片成型领域，涉及一种SERS增强的新冠病毒检测芯片的制备方法及其产品和应用，特别是涉及一种纳米银/纳米氧化锌/纳米金薄膜的制备，及其芯片的成型方法，该芯片可以应用于新冠病毒的检测领域。
背景技术
新型冠状病毒肺炎（COVID-19）疫情的快速蔓延，导致疑似感染者和密切接触者数量激增。尽早进行病毒检测，不仅可以使感染者获得及时治疗，降低死亡风险，还能有效控制传染源，通过隔离切断传播途径。为满足SARS-CoV-2 检测的需求，国内外企业和科研单位陆续研发出众多检测产品。截至2020年3月9日，获批的试剂盒就有15个。目前用于SARS-CoV-2 检测的方法主要有核酸检测法和免疫学检测法，核酸检测法是对病毒RNA 基因组进行检测，包括基因测序、荧光定量PCR、微滴式数字PCR （ddPCR）、基因芯片和环介导等温扩增（LAMP）等技术。免疫学检测法是对病毒抗原或人体免疫反应产生的特异性抗体进行检测，包括免疫色谱试纸条、酶联免疫吸附试验（ELISA）和化学发光免疫分析（CLIA）等技术。核酸检测阳性虽是疑似患者确诊的“金标准”，但其操作繁琐、耗时长，检测结果易受标本质量、病毒感染部位及表达量等众多因素影响，因而核酸单项检测不能满足疫情期间对疑似病例快速筛查的要求。血清特异性抗体是诊断病毒感染的另一关键证据，抗体协同核酸检测可用于辅助诊断和快速筛查。但是如何提高检测的灵敏度/特异度，降低假阴性/假阳性检测结果，仍具有一定的挑战。
发明内容
针对现有技术的不足，本发明目的在于提供一种SERS增强的新冠病毒检测芯片的制备方法。
本发明的另一目的在于：提供一种上述方法制备的SERS增强的新冠病毒检测芯片产品。
本发明的再一目的在于：提供一种上述产品的应用。
本发明的又一目的在于：提供一种上述产品的检测方法。
本发明目的通过下述方案实现：一种SERS增强的新冠病毒检测芯片的制备方法，通过制备纳米银/纳米氧化锌/纳米金的SERS基底，增加荧光分子的发光强度，包括如下步骤：
（1）磁控溅射法镀纳米银薄膜：选用N型硅片Si(111)，分别经过丙酮和乙醇、去离子水、氢氟酸的浸泡与清洗；采用磁控溅射法镀纳米银薄膜，以纯银靶为靶源，真空度3×10-4-5×10-4 Pa，气源为99.99%纯氩气，溅射压强3-6 Pa，沉积速度 0.2-1 Å/s，制得厚度为5-50 nm的纳米银薄膜层的基片；
（2）原子层沉积技术沉积纳米氧化锌：通过原子层沉积技术在步骤（1）所述的纳米银薄膜表面沉积厚度为10-60 nm的纳米氧化锌基片，工艺条件为：将沉积室真空抽至15-20hPa，该基片加热至150-200℃，向沉积室中引入二乙基锌，用高纯氮气清洗沉积室并向沉积室中引入水蒸气，二乙基锌、高纯氮气、水蒸气在沉积室内暴露时间依次为1s、5s、1s、5s；
（3）纳米金颗粒生长：将上述步骤（2）基片浸泡在2-5nm的金颗粒溶液中，振荡2小时后去离子水清洗烘干，进一步浸泡在质量分数为5-25%的碳酸钾溶液中，振荡30分钟后再加入2-10毫升1%高氯酸金溶液，振荡2小时后加入20-200微升巯基乙酸，振荡1-2小时后清洗烘干，得到纳米银/纳米氧化锌/纳米金的芯片；
（4）荧光分子的连接：将制备的纳米银/纳米氧化锌/纳米金的芯片，浸泡在碱性水溶液中，加入等摩尔的1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺（EDC）和N-羟基琥珀酰亚胺（NHS），再加入功能性荧光小分子，常温振荡30-120分钟，洗涤烘干，制备得到SERS增强的新冠病毒检测芯片。
其中，步骤（4）中，所述的碱性水溶液为三乙胺、三乙醇胺、吡啶、尿素中的一种。
在上述方案基础上，步骤（4）中，所述的功能性荧光小分子为异硫氰酸荧光橙红（FITC），罗丹明，磺基氰基-3-马来酰亚胺（cy）、梭基荧光素（FAM）、绿色荧光蛋白（VIC）中的一种。
本发明还提供了一种SERS增强的新冠病毒检测芯片，根据上述方法制备得到。
本发明也提供了一种根据权利要求4所述SERS增强的芯片在新冠病毒检测中的应用。
另外，本发明还提供了一种SERS增强的新冠病毒检测芯片的检测方法，包括下述步骤：
（1）将所述的SERS增强的新冠病毒检测芯片连接采用WHO公布的引物和探针，即OFR 1b基因特异性探针OFR 1b-P的5'端标记FAM，或E基因特异性探针upE-P2的5'端标记VIC，常温振荡30-120分钟；
（2）与新冠病毒的靶物质共培养：靶物质为灭活的新冠病人血样。
本发明芯片应用于新冠病毒的检测中，采用WHO公布的引物和探针，在SERS基底连接荧光探针，实现高灵敏检测，提高检测效率和准确度的同时降低了成本，可用于新型冠状病毒的快速应急检测。
本发明的优点在于：本发明制备的芯片大大增强了小分子的拉曼信号，光学强度高，检测灵敏度高，检测限低。本发明制备过程简单、反应易控制，稳定性好、可产业化，在新冠病毒的检测领域具有广泛的应用前景。
具体实施方式
以下通过具体的实施例对本发明的技术方案作进一步描述。以下的实施例是对本发明的进一步说明，而不限制本发明的范围。
实施例1
一种SERS增强的新冠病毒检测芯片，通过制备纳米银/纳米氧化锌/纳米金的SERS基底，增加荧光分子的发光强度，按如下步骤制备：
（1）磁控溅射法镀纳米银薄膜：选用N型硅片Si(111)，分别经过丙酮和乙醇、去离子水、氢氟酸的浸泡与清洗；采用磁控溅射法镀纳米银薄膜，以纯银靶为靶源，真空度3×10-4Pa，气源为99.99%纯氩气，溅射压强3Pa，沉积速度1 Å/s，制得厚度为5nm的纳米银薄膜层的基片；
（2）原子层沉积技术沉积纳米氧化锌：通过原子层沉积技术在步骤（1）所述的纳米银薄膜表面沉积厚度为10nm的纳米氧化锌，工艺条件为：将沉积室真空抽至15hPa，该基片加热至150℃，向沉积室中引入二乙基锌，用高纯氮气清洗沉积室并向沉积室中引入水蒸气，二乙基锌、高纯氮气、水蒸气在沉积室内暴露时间依次为1s、5s、1s、5s，得到在银薄膜表面沉积厚度为10 nm的纳米氧化锌基片；
（3）纳米金颗粒生长：将上述步骤（2）基片浸泡在2nm的金颗粒溶液中，振荡2小时后去离子水清洗烘干，进一步浸泡在质量分数为5%的碳酸钾溶液中，振荡30分钟后再加入5毫升1%高氯酸金溶液，振荡2小时后加入40微升巯基乙酸，振荡1小时后清洗烘干，得到纳米银/纳米氧化锌/纳米金的芯片；
（4）荧光分子的连接：将制备的纳米银/纳米氧化锌/纳米金的芯片，浸泡在三乙胺水溶液中，加入等摩尔的EDC和NHS，再加入功能性荧光小分子FITC，常温振荡30分钟，洗涤烘干，制备得到一种SERS增强的新冠病毒检测芯片。
芯片检测FITC的光学强度比等量的FITC增加165%。
实施例2
一种SERS增强的新冠病毒检测芯片，与实施例1近似，按如下步骤制备：
（1）磁控溅射法镀纳米银薄膜：选用N型硅片Si(111)，分别经过丙酮和乙醇、去离子水、氢氟酸的浸泡与清洗；采用磁控溅射法镀银薄膜，镀层厚度为25 nm，以纯银靶为靶源，真空度4×10-4 Pa，气源为99.99%纯氩气，溅射压强4Pa，沉积速度 0.5Å/s，制得厚度为25 nm的纳米银薄膜层的基片；
（2）原子层沉积技术沉积纳米氧化锌：通过原子层沉积技术在步骤（1）制备的银薄膜表面沉积厚度为20 nm的纳米氧化锌，其工艺在于：将沉积室真空抽至17 hPa，该基片加热至160℃，向沉积室中引入二乙基锌，用高纯氮气清洗沉积室并向沉积室中引入水蒸气，二乙基锌、高纯氮气、水蒸气在沉积室内暴露时间依次为1s、5s、1s、5s；
（3）纳米金颗粒生长：将上述基片浸泡在4纳米的金颗粒溶液中，振荡2小时后去离子水清洗烘干，进一步浸泡在质量分数为10%的碳酸钾溶液中，振荡30分钟后再加入2毫升1%高氯酸金溶液，振荡2小时后加入100微升巯基乙酸，振荡1小时后清洗烘干，得到纳米银/纳米氧化锌/纳米金的芯片；
（4）荧光分子的连接：将制备的纳米银/纳米氧化锌/纳米金的芯片，浸泡在三乙胺水溶液中，加入等摩尔的EDC和NHS，再加入功能性荧光小分子cy，常温振荡30分钟，洗涤烘干，制备得到SERS增强的新冠病毒检测芯片。
芯片检测cy的光学强度比等量的cy增加214%。
实施例3
一种SERS增强的新冠病毒检测芯片，与实施例1近似，按如下步骤制备：
（1）磁控溅射法镀纳米银薄膜：选用N型硅片Si(111)，分别经过丙酮和乙醇、去离子水、氢氟酸的浸泡与清洗；采用磁控溅射法镀银薄膜，镀层厚度为25 nm，其工艺为：靶源为纯银靶，真空度5×10-4Pa，气源为99.99%纯氩气，溅射压强4Pa，沉积速度 0.5 Å/s，制得厚度为25 nm的纳米银薄膜层的基片；
（2）原子层沉积技术沉积纳米氧化锌：通过原子层沉积技术在步骤（1）所得的银薄膜表面沉积厚度为50 nm的纳米氧化锌，其工艺在于：将沉积室真空抽至20 hPa，基片加热至160℃，向沉积室中引入二乙基锌，用高纯氮气清洗沉积室并向沉积室中引入水蒸气，二乙基锌、高纯氮气、水蒸气在沉积室内暴露时间依次为1s、5s、1s、5s；
（3）纳米金颗粒生长：将上述步骤（2）得到的基片浸泡在2纳米的金颗粒溶液中，振荡2小时后去离子水清洗烘干，进一步浸泡在质量分数为25%的碳酸钾溶液中，振荡30分钟后再加入2毫升1%高氯酸金溶液，振荡2小时后加入100微升巯基乙酸，振荡1小时后清洗烘干，得到纳米银/纳米氧化锌/纳米金的芯片；
（4）将制备的银/氧化锌/金的芯片，浸泡在三乙胺水溶液中，加入等摩尔的EDC和NHS，再加入OFR 1b基因特异性探针，常温振荡30分钟，洗涤烘干，制备得到高SERS效应的芯片可以用于新冠病毒的检测。

Claims (6)

1.一种SERS增强的新冠病毒检测芯片的制备方法，其特征在于，通过制备纳米银/纳米氧化锌/纳米金的SERS基底，增加荧光分子的发光强度，包括如下步骤：
（1）磁控溅射法镀纳米银薄膜：选用N型硅片Si(111)，分别经过丙酮和乙醇、去离子水、氢氟酸的浸泡与清洗；采用磁控溅射法镀纳米银薄膜，以纯银靶为靶源，真空度3×10-4-5×10-4 Pa，气源为99.99%纯氩气，溅射压强3-6 Pa，沉积速度 0.2-1 Å/s，制得厚度为5-50 nm的纳米银薄膜层的基片；
（2）原子层沉积技术沉积纳米氧化锌：通过原子层沉积技术在步骤（1）所述的纳米银薄膜表面沉积厚度为10-60 nm的纳米氧化锌基片，工艺条件为：将沉积室真空抽至15-20hPa，该基片加热至150-200℃，向沉积室中引入二乙基锌，用高纯氮气清洗沉积室并向沉积室中引入水蒸气，二乙基锌、高纯氮气、水蒸气在沉积室内暴露时间依次为1s、5s、1s、5s；
（3）纳米金颗粒生长：将上述步骤（2）基片浸泡在2-5nm的金颗粒溶液中，振荡2小时后去离子水清洗烘干，进一步浸泡在质量分数为5-25%的碳酸钾溶液中，振荡30分钟后再加入2-10毫升1%高氯酸金溶液，振荡2小时后加入20-200微升巯基乙酸，振荡1-2小时后清洗烘干，得到纳米银/纳米氧化锌/纳米金的芯片；
（4）荧光分子的连接：将制备的纳米银/纳米氧化锌/纳米金的芯片，浸泡在碱性水溶液中，加入等摩尔的1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺（EDC）和N-羟基琥珀酰亚胺（NHS），再加入功能性荧光小分子，常温振荡30-120分钟，洗涤烘干，制备得到SERS增强的新冠病毒检测芯片。
2.根据权利要求1所述SERS增强的新冠病毒检测芯片的制备方法，其特征在于，步骤（4）中，所述的碱性水溶液为三乙胺、三乙醇胺、吡啶、尿素中的一种。
3.根据权利要求1或2所述SERS增强的新冠病毒检测芯片的制备方法，其特征在于，步骤（4）中，所述的功能性荧光小分子为异硫氰酸荧光橙红（FITC），罗丹明，磺基氰基-3-马来酰亚胺（cy）、梭基荧光素（FAM）、绿色荧光蛋白（VIC）中的一种。
4.一种SERS增强的新冠病毒检测芯片，其特征在于根据权利要求1-3任一所述方法制备得到。
5.一种根据权利要求4所述SERS增强的芯片在新冠病毒检测中的应用。
6.根据权利要求4所述一种SERS增强的新冠病毒检测芯片的检测方法，其特征在于，包括下述步骤：
（1）将所述的SERS增强的新冠病毒检测芯片连接采用WHO公布的引物和探针，即OFR 1b基因特异性探针OFR 1b-P的5'端标记FAM，或E基因特异性探针upE-P2的5'端标记VIC，常温振荡30-120分钟；
（2）与新冠病毒的靶物质共培养：靶物质为灭活的新冠病人血样。
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112326630A (zh) * 2021-01-04 2021-02-05 中国工程物理研究院激光聚变研究中心 一种用于新型冠状病毒检测的sers芯片及其制备方法
CN113549712A (zh) * 2021-09-22 2021-10-26 广州金域医学检验中心有限公司 基于拉曼光谱的新型冠状病毒核酸检测试剂盒及方法
CN114085930A (zh) * 2022-01-21 2022-02-25 广州国家实验室 用于检测SARS-CoV-2核酸的SERS检测试剂盒及方法
CN114577776A (zh) * 2022-03-01 2022-06-03 哈尔滨工业大学 一种检测液体中新型冠状病毒Spike蛋白的SERS芯片的制备方法和使用方法
CN115142062A (zh) * 2022-05-10 2022-10-04 长春理工大学 一种自清洁复合sers基底及其制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050089901A1 (en) * 2000-09-22 2005-04-28 Porter Marc D. Raman-active reagents and the use thereof
US20100190661A1 (en) * 2009-01-26 2010-07-29 City University Of Hong Kong Sers-active structure for use in raman spectroscopy
CN103837517A (zh) * 2014-03-25 2014-06-04 哈尔滨工业大学 金属薄膜/氧化锌纳米棒阵列荧光增强材料的制备方法
CN104746049A (zh) * 2015-04-07 2015-07-01 南京大学 利用ald制备金属纳米间隙的表面增强拉曼散射基底的方法
CN107543813A (zh) * 2017-08-22 2018-01-05 中国工程物理研究院化工材料研究所 一种表面增强拉曼有序复合阵列芯片的制备方法及其应用
CN109991207A (zh) * 2019-04-25 2019-07-09 福建师范大学 一种用于检测酪氨酸酶的三明治结构的sers传感器及其制备和检测方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050089901A1 (en) * 2000-09-22 2005-04-28 Porter Marc D. Raman-active reagents and the use thereof
US20100190661A1 (en) * 2009-01-26 2010-07-29 City University Of Hong Kong Sers-active structure for use in raman spectroscopy
CN103837517A (zh) * 2014-03-25 2014-06-04 哈尔滨工业大学 金属薄膜/氧化锌纳米棒阵列荧光增强材料的制备方法
CN104746049A (zh) * 2015-04-07 2015-07-01 南京大学 利用ald制备金属纳米间隙的表面增强拉曼散射基底的方法
CN107543813A (zh) * 2017-08-22 2018-01-05 中国工程物理研究院化工材料研究所 一种表面增强拉曼有序复合阵列芯片的制备方法及其应用
CN109991207A (zh) * 2019-04-25 2019-07-09 福建师范大学 一种用于检测酪氨酸酶的三明治结构的sers传感器及其制备和检测方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ANIL KUMAR PAL ET AL.: "Ag/ZnO/Au 3D hybrid structured reusable SERS substrate as highly sensitive platform for DNA detection", 《SENSORS AND ACTUATORS B: CHEMICAL》 *
程昊等: "种子介导法制备SERS基底Ag@Au及对阿莫西林的痕量检测", 《应用化工》 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112326630A (zh) * 2021-01-04 2021-02-05 中国工程物理研究院激光聚变研究中心 一种用于新型冠状病毒检测的sers芯片及其制备方法
CN112326630B (zh) * 2021-01-04 2021-03-16 中国工程物理研究院激光聚变研究中心 一种用于新型冠状病毒检测的sers芯片及其制备方法
CN113549712A (zh) * 2021-09-22 2021-10-26 广州金域医学检验中心有限公司 基于拉曼光谱的新型冠状病毒核酸检测试剂盒及方法
CN113549712B (zh) * 2021-09-22 2022-03-08 广州金域医学检验中心有限公司 基于拉曼光谱的新型冠状病毒核酸检测试剂盒及方法
CN114085930A (zh) * 2022-01-21 2022-02-25 广州国家实验室 用于检测SARS-CoV-2核酸的SERS检测试剂盒及方法
CN114085930B (zh) * 2022-01-21 2022-04-12 广州国家实验室 用于检测SARS-CoV-2核酸的SERS检测试剂盒及方法
CN114577776A (zh) * 2022-03-01 2022-06-03 哈尔滨工业大学 一种检测液体中新型冠状病毒Spike蛋白的SERS芯片的制备方法和使用方法
CN114577776B (zh) * 2022-03-01 2022-10-28 哈尔滨工业大学 一种检测液体中新型冠状病毒Spike蛋白的SERS芯片的制备方法和使用方法
CN115142062A (zh) * 2022-05-10 2022-10-04 长春理工大学 一种自清洁复合sers基底及其制备方法
CN115142062B (zh) * 2022-05-10 2023-10-27 长春理工大学 一种自清洁复合sers基底及其制备方法

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