用于新型冠状病毒检测的纳米材料及生物传感技术

doi:10.15541/jim20220218

无机材料学报 ›› 2023, Vol. 38 ›› Issue (1): 3-31.DOI: 10.15541/jim20220218

• 专栏：抗疫生物材料（特邀编辑: 杨勇） • 上一篇下一篇

用于新型冠状病毒检测的纳米材料及生物传感技术

李妍妍¹^,²(), 彭宇思¹^,², 林成龙¹^,², 罗晓莹³, 滕峥⁴(), 张曦⁴, 黄政仁¹^,², 杨勇¹^,²()

1.中国科学院上海硅酸盐研究所, 高性能陶瓷与超微结构国家重点实验室, 上海 200050
2.中国科学院大学材料科学与光电技术学院, 北京 100049
3.上海交通大学医学院附属仁济医院, 上海市肿瘤研究所癌基因与相关基因国家重点实验室, 上海 200032
4.上海市疾病预防控制中心, 上海 200336

收稿日期:2022-04-12 修回日期:2022-05-03 出版日期:2022-06-22 网络出版日期:2022-06-22
通讯作者: 杨勇, 研究员. E-mail: yangyong@mail.sic.ac.cn;
滕峥, 主任技师. E-mail: tengzheng@scdc.sh.cn
作者简介:李妍妍(1997-), 女, 博士研究生. E-mail: liyanyan20@mails.ucas.ac.cn
基金资助:
国家自然科学基金(52172167);国家重点研发计划国际合作(2021YFE011305)

Nanomaterials and Biosensing Technology for the SARS-CoV-2 Detection

LI Yanyan¹^,²(), PENG Yusi¹^,², LIN Chenglong¹^,², LUO Xiaoying³, TENG Zheng⁴(), ZHANG Xi⁴, HUANG Zhengren¹^,², YANG Yong¹^,²()

1. State Key Laboratory of High Performance Ceramics and Superfine Microstructures, Shanghai Institute of Ceramics, Chinese Academy of Sciences, Shanghai 200050, China
2. College of Materials Science and Opto-Electronics Engineering, University of Chinese Academy of Sciences, Beijing 100049, China
3. State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200032, China
4. Shanghai Provincial Center for Disease Control and Prevention, Shanghai 200336, China

Received:2022-04-12 Revised:2022-05-03 Published:2022-06-22 Online:2022-06-22
Contact: YANG Yong, professor. E-mail: yangyong@mail.sic.ac.cn;
TENG Zheng, professor. E-mail: tengzheng@scdc.sh.cn
About author:LI Yanyan (1997-), female, PhD candidate. E-mail: liyanyan20@mails.ucas.ac.cn
Supported by:
National Natural Science Foundation of China(52172167);National Key R&D Program of China(2021YFE011305)

摘要/Abstract

摘要：

新型冠状病毒肺炎(Corona Virus Disease 2019, COVID-19)疫情大流行引起全球对此重大突发公共卫生事件的高度关注。新型冠状病毒(SARS-CoV-2)经过多次突变, 出现传染速度加快、免疫逃逸、隐匿性传播等特性, 令防控形势至今仍异常严峻。对患者的早发现、早隔离仍然是目前最有效的防控措施。因此, 迫切需要快速、高灵敏的检测手段来甄别此病毒, 以便及早识别感染者。本文简要介绍了SARS-CoV-2的一般特征, 并针对核酸、抗体、抗原及病原体作为检测靶标的不同检测手段及最新进展进行分类概述; 对一些光学、电学、磁学以及可视化的新型纳米传感器在SARS-CoV-2检测技术上的应用进行了分析。鉴于纳米技术的应用在提高检测灵敏度、特异性以及准确率上具有优势, 本文详细介绍了新型纳米传感器在SARS-CoV-2检测中的研究进展, 包括表面增强拉曼基生物传感器、电化学生物传感器、磁纳米生物传感器以及比色生物传感器等, 并探讨了纳米材料在新型生物传感器构建中的作用和挑战, 为纳米材料研究人员开发各种类型的冠状病毒传感技术提供思路。

关键词: SARS-CoV-2, 检测方法, 核酸, 抗体, 抗原, 纳米材料, 生物传感器, 综述

Abstract:

The outbreak of corona virus disease 2019 (COVID-19) has aroused great attention around the world. SARS-CoV-2 possesses characteristics of faster transmission, immune escape, and occult transmission by many mutation, which caused still grim situation of prevention and control. Early detection and isolation of patients are still the most effective measures at present. So, there is an urgent need for new rapid and highly sensitive testing tools to quickly identify infected patients as soon as possible. This review briefly introduces general characteristics of SARS-CoV-2, and provides recentl overview and analysis based on different detection methods for nucleic acids, antibodies, antigens as detection target. Novel nano-biosensors for SARS-CoV-2 detection are analyzed based on optics, electricity, magnetism, and visualization. In view of the advantages of nanotechnology in improving detection sensitivity, specificity and accuracy, the research progress of new nano-biosensors is introduced in detail, including SERS-based biosensors, electrochemical biosensors, magnetic nano-biosensors and colorimetric biosensors. Functions and challenges of nano-materials in construction of new nano-biosensors are discussed, which provides ideas for the development of various coronavirus biosensing technologies for nanomaterial researchers.

Key words: SARS-CoV-2, detection method, nucleic acid, antibody, antigen, nanomaterial, biosensor, review

中图分类号:

TQ174

李妍妍, 彭宇思, 林成龙, 罗晓莹, 滕峥, 张曦, 黄政仁, 杨勇. 用于新型冠状病毒检测的纳米材料及生物传感技术[J]. 无机材料学报, 2023, 38(1): 3-31.

LI Yanyan, PENG Yusi, LIN Chenglong, LUO Xiaoying, TENG Zheng, ZHANG Xi, HUANG Zhengren, YANG Yong. Nanomaterials and Biosensing Technology for the SARS-CoV-2 Detection[J]. Journal of Inorganic Materials, 2023, 38(1): 3-31.

图/表 20

图1 SARS-CoV-2的TEM照片(A)和示意图(B)[9]

Fig. 1 TEM image (A) and schemetic morphology (B) of SARS-CoV-2[9] The color figures can be obtained from online edition

图2 冠状病毒的分类(a、c)、感染症状(b)及其结构示意图(d)[16]

Fig. 2 Classification (a, c), symptoms (b) and structure (c) of coronavirus[16] The color figures can be obtained from online edition

图3 用于检测SARS-CoV-2的微流控基因传感器[35]

Fig. 3 Scheme of the lab-on-a-chip genosensor for SARS- CoV-2 virus detection[35] WE: working electrode; CE: counter electrode; RE: reference electrode; ssDNA: single strand DNA The color figure can be obtained from online edition

图4 ddPCR在临床样本分析中的基本工作流程[37]

Fig. 4 Principle workflow of ddPCR in analyzing clinical samples[37] The color figure can be obtained from online edition

图5 基于CRISPR-Cas12侧向流动技术检测SARS-CoV-2[44]

Fig. 5 Detection of SARS-CoV-2 based on CRISPR-Cas12 combined with lateral flow technique[44] The color figure can be obtained from online edition

图6 病毒感染后人体免疫反应变化[16]

Fig. 6 Change of human immune response after viral infection[16] The color figure can be obtained from online edition

图7 检测SARS-CoV-2的ELISA技术[16]

Fig. 7 ELISA method for detection of SARS-CoV-2[16] ACE2: angiotensin converting enzyme 2; HRP: horse radish peroxidase; ELISA: enzyme linked immunosorbent assay The color figure can be obtained from online edition

表1 常规SARS-CoV-2检测方法比较

Table 1 Comparison of conventional detection methods for SARS-CoV-2

Object sample	Characteristic	Detection technology	Advantage	Disadvantage
RNA	1. Target: the gene sequences of SARS-CoV-2 2. Great producibility 3. Long detection period 4. Possibility of being contaminated and false positive result	Whole genome sequencing	1. High accuracy and sensitivity 2. Reflecting genetic information of pathogen comprehensively	1. Expensive special instruments 2. Relying on professionals 3. Difficulty in detection on a large scale
		RT-qPCR	1. High sensitivity and specificity 2. Low cost	1. Long amplification time 2. High requirements of equipment 3. Complex operation
		LAMP	1. Isothermal reaction 2. High efficiency and speed 3. High sensitivity and visualization	1. Complex design of primer 2. Low specificity
		Microfluidic chip	1. Multiple detection of pathogens 2. Integration of sample preparation and detection 3. Ability in automate analysis	Difficulty in chip design, material selection, processing, packaging, and storage
		ddPCR	1. High sensitivity and lowest limitation of detection 2. Facilitation and high degree of automation 3. Quantitative detection	1. Small reaction volume 2. Expensive equipment and reagents
		CRISPR	1. High speed and low cost 2. High sensitivity 3. Strong system stability 4. On-site detection	The accuracy of detection needs to be verified
Antibodies	1. Target: human antibodies stimulated by SARS-CoV-2 2. Easy sample collection and low detection threshold 3. Simple operation and high throughput 4. Limitation of timeframe 5. Lower sensitivity and specificity than those of nucleic acid detection	ELISA	1. Low difficulty of standardization of carrier 2. High sensitivity and specificity 3. Simple equipment	1. Long detection time and cumbersome steps 2. Limited single detection throughout
		LFIA (Colloidal gold method)	1. On-site detection caused by easy operation 2. High sensitivity and speed 3. Low cost 4. Mass production	1. Only qualitative analysis 2. Different reproducibility of different batches of products
		CLIA	1. High sensitivity and specificity 2. High throughput detection and high degree of automation	1. Special instrument 2. High detection cost
Antigen	1. Target: SARS-CoV-2 antigen 2. Simple and fast operation	LFIA (Colloidal gold method)	1. Fast and facile operation 2. Visualization 3. On-site detection and large-scale population screening	Low sensitivity

表1 常规SARS-CoV-2检测方法比较

Table 1 Comparison of conventional detection methods for SARS-CoV-2

Object sample	Characteristic	Detection technology	Advantage	Disadvantage
RNA	1. Target: the gene sequences of SARS-CoV-2 2. Great producibility 3. Long detection period 4. Possibility of being contaminated and false positive result	Whole genome sequencing	1. High accuracy and sensitivity 2. Reflecting genetic information of pathogen comprehensively	1. Expensive special instruments 2. Relying on professionals 3. Difficulty in detection on a large scale
		RT-qPCR	1. High sensitivity and specificity 2. Low cost	1. Long amplification time 2. High requirements of equipment 3. Complex operation
		LAMP	1. Isothermal reaction 2. High efficiency and speed 3. High sensitivity and visualization	1. Complex design of primer 2. Low specificity
		Microfluidic chip	1. Multiple detection of pathogens 2. Integration of sample preparation and detection 3. Ability in automate analysis	Difficulty in chip design, material selection, processing, packaging, and storage
		ddPCR	1. High sensitivity and lowest limitation of detection 2. Facilitation and high degree of automation 3. Quantitative detection	1. Small reaction volume 2. Expensive equipment and reagents
		CRISPR	1. High speed and low cost 2. High sensitivity 3. Strong system stability 4. On-site detection	The accuracy of detection needs to be verified
Antibodies	1. Target: human antibodies stimulated by SARS-CoV-2 2. Easy sample collection and low detection threshold 3. Simple operation and high throughput 4. Limitation of timeframe 5. Lower sensitivity and specificity than those of nucleic acid detection	ELISA	1. Low difficulty of standardization of carrier 2. High sensitivity and specificity 3. Simple equipment	1. Long detection time and cumbersome steps 2. Limited single detection throughout
		LFIA (Colloidal gold method)	1. On-site detection caused by easy operation 2. High sensitivity and speed 3. Low cost 4. Mass production	1. Only qualitative analysis 2. Different reproducibility of different batches of products
		CLIA	1. High sensitivity and specificity 2. High throughput detection and high degree of automation	1. Special instrument 2. High detection cost
Antigen	1. Target: SARS-CoV-2 antigen 2. Simple and fast operation	LFIA (Colloidal gold method)	1. Fast and facile operation 2. Visualization 3. On-site detection and large-scale population screening	Low sensitivity

图8 SERS基免疫平台示意图[77]

Fig. 8 Schematic illustration of the SERS-based immunoassay[77] MBA: thiosalicylic acid; BSA: bovine serum albumin; NPs: nanoparticles The color figure can be obtained from online edition

图9 SERS生物传感器精准捕获待检SARS-CoV-2病毒示意图[8]

Fig. 9 Schematic diagram of operation procedure of COVID-19 SERS sensor[8] The color figure can be obtained from online edition

图10 基于SnS2微球的两步SERS检测法诊断SARS-CoV-2活性及其传染性[80]

Fig. 10 Application of SnS2 microspheres for diagnosing the infectiousness of SARS-CoV-2[80] (A) Experimental procedure for diagnosing the infectiousness of SARS-CoV-2; (B) SVM analysis results to identify the mixture of the SARS-CoV-2 with complete viral structure and the lysed SARS-CoV-2; (C) Raman scattering diagram of three contamination situations of the novel coronavirus based on SnS2 substrates; (D) SVM analysis results to identify the lysed SARS-CoV-2; (E) SVM analysis results to identify the mixture of the SARS-CoV-2 with complete viral structure and the lysed SARS-CoV-2 after eliminating RNA and relysing virus samples; (F) SVM analysis results to identify the SARS-CoV-2 with complete viral structure; (G) SVM analysis results to identify the lysed SARS-CoV-2 after eliminating RNA and relysing virus samples. SVM: support vector machine The color figure can be obtained from online edition

图11 纳米等离子光学传感芯片检测SARS-CoV-2颗粒示意图[89]

Fig. 11 Schematic diagram of nano-plasma optic sensor for detection of SARS-CoV-2[89] (A) Schematic diagram of the nanoplasmonic resonance sensor for determination of SARS-CoV-2 pseudovirus concentration; (B) Photograph (middle) of one piece of Au nanocup array chip with a drop of water on top The color figure can be obtained from online edition

图12 基于5G支持的荧光生物传感器用于SARS-CoV-2检测[100]

Fig. 12 SARS-CoV-2 detection based on 5G-enabled fluorescence biosensor[100] (a) The principle of the UCNPs based lateral flow assay in detection of SARS-CoV-2; (b) The working process of the proposed 5G-enabled fluorescence sensor; (c) The circuit configuration and hardware composition of the fluorescence sensor; CL: control line; TL1: test line 1; TL2: test line 2; UCNPs: up-conversion nanoparticles; EEPROM: electrically erasable programmable read only memory; ADC: application data center; MCU: motor control unit The color figure can be obtained from online edition

图13 用于检测未处理唾液中SARS-CoV-2的磁珠基电化学分析平台[126]

Fig. 13 Magnetic beads-based electrochemical assay for SARS-CoV-2 detection in untreated saliva[126] MBs: magnetic beads; MAb: monoclonal antibody; PAb: polyclonal antibody; AP: alkaline phosphatase; CB-SPE: carbon-based screen-printed electrodes The color figure can be obtained from online edition

图14 基于电化学生物传感器分析SARS-CoV-2 RNA的检测原理[136]

Fig. 14 Principle of the proposed electrochemical biosensor for sensitive analysis of SARS-CoV-2 RNA[136] HP: hairpin; TdT: terminal deoxynucleotidyl transferase; dNTP: deoxyribonucleotides; DPV: differential pulse voltammetry The color figure can be obtained from online edition

图15 基于PMO修饰的G-FET纳米传感器快速直接识别SARS-CoV-2的示意图[148]

Fig. 15 Schematic diagram of rapid direct identification of SARS-CoV-2 using PMO-functionalized G-FET nano-sensors[148] G-FET: graphene field-effect transistor; PMO: phosphorodiamidate morpholino oligos The color figure can be obtained from online edition

图16 基于磁颗粒光谱生物传感器的SARS-CoV-2核酸检测示意图[166]

Fig. 16 Schematic diagram of detection of SARS-CoV-2 RNA based on magnetic particle spectroscopy biosensors[166] (A) Magnetic nanoparticles (gold) and polystyrene beads (silver) with streptavidin (purple)-modified surface are equipped with single stranded DNA strands (red and green, respectively) with a specific sequence via biotin-streptavidin-binding; (B) Applying a sinusoidal magnetic field (black) to a solution of nanoparticles results in reorientation of the nanoparticles which can be readout by measuring the magnetic response M of the nanoparticles; (C) Exemplary spectrum of the ratio of received harmonics as a function of excitation frequencies for 80 nm BNF magnetic particles The color figure can be obtained from online edition

图17 基于超低场核磁共振的磁弛豫开关分析的SARS-CoV-2检测过程[169]

Fig. 17 Detection process of SARS-CoV-2 of the magnetic relaxation switches assay with ULF NMR[169] ULF: ultra-low field; GPG: Gd3+ loaded PEG modified GQDs; GQDs: graphene quantum dots The color figure can be obtained from online edition

图18 基于比色法的SARS-CoV-2检测示意图[187]

Fig. 18 Schematic diagram of the detection of SARS-CoV-2 based on colorimetric biosensors[187] UTM: universal transport medium The color figure can be obtained from online edition

表2 用于SARS-CoV-2检测的新型生物传感器比较

Table 2 Comparison of novel biosensors for SARS-CoV-2 detection

Detection technology	Detection method	Object	Sample	Related material	Detection time	Lower detection limit	Ref.
SERS-based biosensors	Labelled-SERS	S protein	Lysis solution	Macro/nanostructure Au substrate	15 min	10 PFU/mL	[73]
SERS-based biosensors	Label-free SERS	Virus particles	Nasal/throat solution	Macro/nanostructure Au substrate, Au nanoparticles	15 min	60 copies/mL	[72]
SPR-based biosensors	Combining SPR and LSPR	Pseudovirus particles	N/A	Macro/nanostructure Au substrate, Au nanoparticles	15 min	370 vp/mL	[89]
Fluorescence biosensors	“signal on” mode	RNA	Lysis solution	N/A	15 samples/ 45 min	600 copies/mL	[102]
Electrochemical biosensors	Voltammetric/ amperometric biosensors	RNA	Nasal/throat solution	Au nanoparticles	5 min	6900 copies/mL	[128]
	Impedimetric biosensors	Antibodies	Serum	Au nanoparticles	30 min	N/A	[135]
	Potentiometric biosensors	Cholinesterase	Blood	Graphene and copper	~7 s (only detection time)	7.9 × 10^-8mol/L	[139]
	FET-based biosensors	RNA	Nasal/throat solution	Graphene	1 min (only detection time)	10-20 copies/mL	[150]
Magnetic biosensors	Magnetoresistance	Antibodies	Blood	Magnetic nanoparticles	10 min	5-10 ng/mL	[162]
	Magnetic particle spectroscopy platforms	S protein and N protein	PBS	Magnetic nanoparticles	N/A	1.56 nmol/L	[165]
	Nuclear magnetic resonance	Antibodies	Blood	Magnetic graphene quantum dot	2 min	248 vp/mL	[169]
Colorimetric biosensors	Agglomeration of nanoparticles	RNA	N/A	Au nanoparticles	>45 min	160 fmol/L	[185]

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用于新型冠状病毒检测的纳米材料及生物传感技术

Nanomaterials and Biosensing Technology for the SARS-CoV-2 Detection

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