SARS-CoV-2抗原快速检测方法的研究进展

doi:10.3969/j.issn.1673-8640.2023.01.019

摘要/Abstract

摘要：

新型冠状病毒（SARS-CoV-2）感染的大流行导致了一场全球性的健康危机。快速、准确地检出SARS-CoV-2是明确感染规模并采取进一步干预措施的重要前提。病毒核酸检测是新型冠状病毒肺炎（COVID-19）确诊的标准方法，在一些特殊场景下，即时检验（POCT）是必要的补充。文章对用于SARS-CoV-2抗原检测的POCT方法的标记物（胶体金、乳胶微球、荧光物质、量子点和酶）和基于不同方法构建的各种类型的传感器进行总结，以期为SARS-CoV-2检测试剂的研发提供参考。

关键词: 新型冠状病毒, 新型冠状病毒肺炎, 即时检验, 抗原检测

Abstract:

Severe acute respiratory syndrome coronavirus 2（SARS-CoV-2）infection has led to a global health crisis. Rapid and accurate determination of SARS-CoV-2 is an important prerequisite for determining the scale of infection and further taking medical interventions or government actions. Although nucleic acid test is still the standard method for diagnosing corona virus disease 2019（COVID-19），antigen test methods that are easier to achieve point-of-care testing（POCT）are still indispensable additions under some conditions. This review mainly summarizes the research progress of SARS-CoV-2 antigen test POCT methods，such as colloidal gold，latex microspheres，fluorescent material，quantum dot and enzyme，and briefly introduces various types of sensors based on different methods，in order to provide a reference for the development of SARS-CoV-2 diagnostic reagents.

Key words: Severe acute respiratory syndrome coronavirus 2, Corona virus disease 2019, Point-of-care testing, Antigen test

中图分类号:

R373.1

岳彩蝶, 李军燕, 丁爱军, 谢力, 曾韦锟. SARS-CoV-2抗原快速检测方法的研究进展[J]. 检验医学, 2023, 38(1): 87-93.

YUE Caidie, LI Junyan, DING Aijun, XIE Li, ZENG Weikun. Research progress of rapid determination methods for SARS-CoV-2 antigen[J]. Laboratory Medicine, 2023, 38(1): 87-93.

图/表 1

参考文献 37

[1]	World Health Organization. WHO coronavirus(COVID-19) dashboard[EB/OL]. (2022-04-28)[2022-04-29]. https://covid19.who.int/.
[2]	World Health Organization. Antigen-detection in the diagnosis of SARS-CoV-2 infection[EB/OL]. (2021-10-06)[2021-10-06]. https://www.who.int/publications/i/item/antigen-detection-in-the-diagnosis-of-sars-cov-2infection-using-rapid-immunoassays.
[3]	FDA. At-home OTC COVID-19 diagnostic tests[EB/OL]. (2022-04-27)[2022-04-28]. https://www.fda.gov/medical-devices/coronavirus-covid-19-and-medical-devices/home-otc-covid-19-diagnostic-tests?utm_medium=email&utm_source=govdelivery#list.
[4]	WALLS A C, PARK Y J, TORTORICI M A, et al. Structure,function,and antigenicity of the SARS-CoV-2 spike glycoprotein[J]. Cell, 2020, 181(2):281-292.e6. DOI URL
[5]	DRAIN P K, AMPAJWALA M, CHAPPEL C, et al. A rapid,high-sensitivity SARS-CoV-2 nucleocapsid immunoassay to aid diagnosis of acute COVID-19 at the point of care:a clinical performance study[J]. Infect Dis Ther, 2021, 10(2):753-761. DOI
[6]	PAULES C I, MARSTON H D, FAUCI A S. Coronavirus infections-more than just the common cold[J]. JAMA, 2020, 323(8):707-708. DOI PMID
[7]	国家药品监督管理局. 国家药监局已批准31个新冠病毒抗原检测试剂[EB/OL]. (2022-04-29)[2022-04-29]. https://www.nmpa.gov.cn/yaowen/ypjgyw/20220429161546149.html.
[8]	EU. Common list of COVID-19 rapid antigen tests that member states mutually recognise[EB/OL]. (2022-04-08)[2022-04-08]. https://european-union.europa.eu/index_en?wt-search=yes.
[9]	LI G, WANG A, CHEN Y, et al. Development of a colloidal gold-based immunochromatographic strip for rapid detection of severe acute respiratory syndrome coronavirus 2 spike protein[J]. Front Immunol, 2021, 12:635677. DOI URL
[10]	PENG T, JIAO X, LIANG Z, et al. Lateral flow immunoassay coupled with copper enhancement for rapid and sensitive SARS-CoV-2 nucleocapsid protein detection[J]. Biosensors(Basel), 2021, 12(1):13.
[11]	CHEN X, KANG S, IKBAL M A, et al. Synthetic nanobody-functionalized nanoparticles for accelerated development of rapid,accessible detection of viral antigens[J]. Biosens Bioelectron, 2022, 202:113971. DOI URL
[12]	LIU Y, ZHAN L, SHEN J W, et al. fM-aM detection of the SARS-CoV-2 antigen by advanced lateral flow immunoassay based on gold nanospheres[J]. ACS Appl Nano Mater, 2021, 4(12):13826-13837. DOI PMID
[13]	丽珠医学. 助力抗疫——丽珠集团新冠抗原检测试剂获批上市[EB/OL]. (2022-04-13)[2022-04-13]. https://www.livzon.com.cn/news/738.html.
[14]	SHEN L, ZHANG Q, LUO X, et al. A rapid lateral flow immunoassay strip for detection of SARS-CoV-2 antigen using latex microspheres[J]. J Clin Lab Anal, 2021, 35(12):e24091.
[15]	RUSANEN J, KAREINEN L, SZIROVICZA L, et al. A generic,scalable,and rapid time-resolved förster resonance energy transfer-based assay for antigen detection-SARS-CoV-2 as a proof of concept[J]. mBio, 2021, 12(3):e00902-21.
[16]	DIAO B, WEN K, ZHANG J, et al. Accuracy of a nucleocapsid protein antigen rapid test in the diagnosis of SARS-CoV-2 infection[J]. Clin Microbiol Infect, 2021, 27(2):289.e1-289.e4.
[17]	WU K, CHUGH V K, D KRISHNA V, et al. One-step,wash-free,nanoparticle clustering-based magnetic particle spectroscopy bioassay method for detection of SARS-CoV-2 spike and nucleocapsid proteins in the liquid phase[J]. ACS Appl Mater Interfaces, 2021, 13(37):44136-44146. DOI URL
[18]	LIU J, RUAN G, MA W, et al. Horseradish peroxidase-triggered direct in situ fluorescent immunoassay platform for sensing cardiac troponin I and SARS-CoV-2 nucleocapsid protein in serum[J]. Biosens Bioelectron, 2022, 198:113823. DOI URL
[19]	LIU D, JU C, HAN C, et al. Nanozyme chemiluminescence paper test for rapid and sensitive detection of SARS-CoV-2 antigen[J]. Biosens Bioelectron, 2020, 173:112817. DOI URL
[20]	WANG C, WANG C, QIU J, et al. Ultrasensitive,high-throughput,and rapid simultaneous detection of SARS-CoV-2 antigens and IgG/IgM antibodies within 10 min through an immunoassay biochip[J]. Mikrochim Acta, 2021, 188(8):262. DOI
[21]	EISSA S, ZOUROB M. Development of a low-cost cotton-tipped electrochemical immunosensor for the detection of SARS-CoV-2[J]. Anal Chem, 2021, 93(3):1826-1833. DOI PMID
[22]	MEHMANDOUST M, GUMUS Z P, SOYLAK M, et al. Electrochemical immunosensor for rapid and highly sensitive detection of SARS-CoV-2 antigen in the nasal sample[J]. Talanta, 2022, 240:123211. DOI URL
[23]	IDILI A, PAROLO C, ALVAREZ-DIDUK R, et al. Rapid and efficient detection of the SARS-CoV-2 spike protein using an electrochemical aptamer-based sensor[J]. ACS Sens, 2021, 6(8):3093-3101. DOI URL
[24]	RAZIQ A, KIDAKOVA A, BOROZNJAK R, et al. Development of a portable MIP-based electrochemical sensor for detection of SARS-CoV-2 antigen[J]. Biosens Bioelectron, 2021, 178:113029. DOI URL
[25]	LIV L, ÇOBAN G, NAKIBOLU N, et al. A rapid,ultrasensitive voltammetric biosensor for determining SARS-CoV-2 spike protein in real samples[J]. Biosens Bioelectron, 2021, 192:113497. DOI URL
[26]	HASHEMI S A, GOLAB BEHBAHAN N G, BAHRANI S, et al. Ultra-sensitive viral glycoprotein detection NanoSystem toward accurate tracing SARS-CoV-2 in biological/non-biological media[J]. Biosens Bioelectron, 2021, 171:112731. DOI URL
[27]	SEO G, LEE G, KIM M J, et al. Rapid detection of COVID-19 causative virus(SARS-CoV-2) in human nasopharyngeal swab specimens using field-effect transistor-based biosensor[J]. ACS Nano, 2020, 14(4):5135-5142. DOI URL
[28]	GAO J, WANG C, CHU Y, et al. Graphene oxide-graphene Van der Waals heterostructure transistor biosensor for SARS-CoV-2 protein detection[J]. Talanta, 2022, 240:123197. DOI URL
[29]	CHO S Y, JIN X, GONG X, et al. Antibody-free rapid detection of SARS-CoV-2 proteins using corona phase molecular recognition to accelerate development time[J]. Anal Chem, 2021, 93(44):14685-14693. DOI URL
[30]	KIM H Y, LEE J H, KIM M J, et al. Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins[J]. Biosens Bioelectron, 2021, 175:112868. DOI URL
[31]	GUO J, CHEN S, TIAN S, et al. 5G-enabled ultra-sensitive fluorescence sensor for proactive prognosis of COVID-19[J]. Biosens Bioelectron, 2021, 181:113160. DOI URL
[32]	WANG C, CHENG X, LIU L, et al. Ultrasensitive and simultaneous detection of two specific SARS-CoV-2 antigens in human specimens using direct/enrichment dual-mode fluorescence lateral flow immunoassay[J]. ACS Appl Mater Interfaces, 2021, 13(34):40342-40353. DOI URL
[33]	GEORGAS A, LAMPAS E, HOUHOULA D P, et al. ACE2-based capacitance sensor for rapid native SARS-CoV-2 detection in biological fluids and its correlation with real-time PCR[J]. Biosens Bioelectron, 2022, 202:114021. DOI URL
[34]	QI H, HU Z, YANG Z, et al. Capacitive aptasensor coupled with microfluidic enrichment for real-time detection of trace SARS-CoV-2 nucleocapsid protein[J]. Anal Chem, 2022, 94(6):2812-2819. DOI PMID
[35]	HUANG L, DING L, ZHOU J, et al. One-step rapid quantification of SARS-CoV-2 virus particles via low-cost nanoplasmonic sensors in generic microplate reader and point-of-care device[J]. Biosens Bioelectron, 2021, 171:112685. DOI URL
[36]	DAI Z, XU X, WANG Y, et al. Surface plasmon resonance biosensor with laser heterodyne feedback for highly-sensitive and rapid detection of COVID-19 spike antigen[J]. Biosens Bioelectron, 2022, 206:114163. DOI URL
[37]	FORINOVÁ M, PILIPENCO A, VÍŠOVÁ I, et al. Functionalized terpolymer-brush-based biointerface with improved antifouling properties for ultra-sensitive direct detection of virus in crude clinical samples[J]. ACS Appl Mater Interfaces, 2021, 13(50):60612-60624. DOI URL

传感器类型	研究者	基本原型	靶标	检测优势
电容式生物传感器	GEORGAS等^[33]	将ACE2蛋白固定在金叉指电极表面上的无抗体生物传感器	S蛋白抗原	检测时间为2 min；可区分临床阳性样本（7份）和阴性样本（16份）；检测限为750 pg/（μL·mm²）；复现性>95%
	QI等^[34]	与微电极阵列芯片结合的定量电容式适配体传感器	N蛋白抗原	检测时间为15 s；线性范围为10^-5~10^-2 ng/mL；无标记、实时、易操作；对目标抗原的选择性高达6 369∶1；临床样本（9份阳性、9份阴性）验证均达到预期结果
光学生物传感器	HUANG等^[35]	病毒S蛋白特异性纳米共振传感器	S蛋白抗原	检测时间为15 min；检测限为370 VP/mL；可在酶标仪和移动设备上显示结果；低成本、高通量
	DAI等^[36]	基于激光外差反馈的新型表面等离子体共振生物传感器	S蛋白抗原	检测时间为1 min；线性范围为0.01~1 000 ng/mL；检测限为0.08 pg/mL
压电生物传感器	FORINOVá等^[37]	由抗污染三元共聚物生物接口改良的压电生物传感器	N蛋白抗原	检测时间为20 min；检测限为1.3×10⁴ PFU/mL；经质谱和RT-qPCR验证，特异性高；抗污染能力强

传感器类型	研究者	基本原型	靶标	检测优势
电容式生物传感器	GEORGAS等^[33]	将ACE2蛋白固定在金叉指电极表面上的无抗体生物传感器	S蛋白抗原	检测时间为2 min；可区分临床阳性样本（7份）和阴性样本（16份）；检测限为750 pg/（μL·mm²）；复现性>95%
	QI等^[34]	与微电极阵列芯片结合的定量电容式适配体传感器	N蛋白抗原	检测时间为15 s；线性范围为10^-5~10^-2 ng/mL；无标记、实时、易操作；对目标抗原的选择性高达6 369∶1；临床样本（9份阳性、9份阴性）验证均达到预期结果
光学生物传感器	HUANG等^[35]	病毒S蛋白特异性纳米共振传感器	S蛋白抗原	检测时间为15 min；检测限为370 VP/mL；可在酶标仪和移动设备上显示结果；低成本、高通量
	DAI等^[36]	基于激光外差反馈的新型表面等离子体共振生物传感器	S蛋白抗原	检测时间为1 min；线性范围为0.01~1 000 ng/mL；检测限为0.08 pg/mL
压电生物传感器	FORINOVá等^[37]	由抗污染三元共聚物生物接口改良的压电生物传感器	N蛋白抗原	检测时间为20 min；检测限为1.3×10⁴ PFU/mL；经质谱和RT-qPCR验证，特异性高；抗污染能力强