The development of a highly sensitive and quantitative SARS-CoV-2 rapid antigen test applying newly developed monoclonal antibodies to an automated chemiluminescent flow-through membrane immunoassay device.

IF 2.9 4区 医学 Q3 IMMUNOLOGY BMC Immunology Pub Date : 2023-09-26 DOI:10.1186/s12865-023-00567-y
Kengo Nishimura, Hiroaki Kitazawa, Takashi Kawahata, Kosuke Yuhara, Takahiro Masuya, Toshihiro Kuroita, Kentarou Waki, Seiichi Koike, Masaharu Isobe, Nobuyuki Kurosawa
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Abstract

Background: Rapid and accurate diagnosis of individuals with SARS-CoV-2 infection is an effective way to prevent and control the spread of COVID-19. Although the detection of SARS-CoV-2 viral RNA by RT-qPCR is the gold standard for COVID-19 testing, the use of antigen-detecting rapid diagnostic tests (Ag-RDTs) is emerging as a complementary surveillance tool as Omicron case numbers skyrocket worldwide. However, the results from Ag-RDTs are less accurate in individuals with low viral loads.

Results: To develop a highly sensitive and accurate Ag-RDT, 90 monoclonal antibodies were raised from guinea pigs immunized with SARS CoV-2 nucleocapsid protein (CoV-2-NP). By applying a capture antibody recognizing the structural epitope of the N-terminal domain of CoV-2-NP and a detection antibody recognizing the C-terminal tail of CoV-2-NP to an automated chemiluminescence flow-through membrane immunoassay device, we developed a novel Ag-RDT, CoV-2-POCube. The CoV-2-POCube exclusively recognizes CoV-2-NP variants but not the nucleocapsid proteins of other human coronaviruses. The CoV-2-POCube achieved a limit of detection sensitivity of 0.20 ~ 0.66 pg/mL of CoV-2-NPs, demonstrating more than 100 times greater sensitivity than commercially available SARS-CoV-2 Ag-RDTs.

Conclusions: CoV-2-POCube has high analytical sensitivity and can detect SARS-CoV-2 variants in 15 min without observing the high-dose hook effect, thus meeting the need for early SARS-CoV-2 diagnosis with lower viral load. CoV-2-POCube is a promising alternative to currently available diagnostic devices for faster clinical decision making in individuals with suspected COVID-19 in resource-limited settings.

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将新开发的单克隆抗体应用于自动化学发光流通膜免疫测定装置的高度灵敏和定量的严重急性呼吸系统综合征冠状病毒2型快速抗原检测的开发。
背景:快速准确地诊断SARS-CoV-2感染者是预防和控制新冠肺炎传播的有效途径。尽管RT-qPCR检测SARS-CoV-2病毒RNA是新冠肺炎检测的金标准,但随着全球奥密克戎病例数飙升,抗原检测快速诊断检测(Ag-RDT)的使用正在成为一种补充监测工具。然而,Ag-RDT的结果在病毒载量低的个体中不太准确。结果:从SARS冠状病毒2型核衣壳蛋白(CoV-2-NP)免疫的豚鼠身上制备了90株单克隆抗体,建立了一种高灵敏、准确的Ag-RDT。通过将识别CoV-2-NP N端结构域结构表位的捕获抗体和识别CoV-2-NP C端尾部的检测抗体应用于自动化学发光流通膜免疫测定装置,我们开发了一种新型的Ag-RDT,CoV-2-POCube。CoV-2-POCube只识别CoV-2-NP变体,但不识别其他人类冠状病毒的核衣壳蛋白。CoV-2-POCube实现了0.20的检测灵敏度极限 ~ 0.66 pg/mL的CoV-2-NP,显示出比市售的严重急性呼吸系统综合征冠状病毒2型Ag-RDT高100多倍的灵敏度。CoV-2-POCube是目前可用诊断设备的一种很有前途的替代品,可以在资源有限的环境中为疑似新冠肺炎患者更快地做出临床决策。
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来源期刊
BMC Immunology
BMC Immunology 医学-免疫学
CiteScore
5.50
自引率
0.00%
发文量
54
审稿时长
1 months
期刊介绍: BMC Immunology is an open access journal publishing original peer-reviewed research articles in molecular, cellular, tissue-level, organismal, functional, and developmental aspects of the immune system as well as clinical studies and animal models of human diseases.
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