Automated, flow-based chemiluminescence microarray immunoassay for the rapid multiplex detection of IgG antibodies to SARS-CoV-2 in human serum and plasma (CoVRapid CL-MIA)

Abstract

In the face of the COVID-19 pandemic, the need for rapid serological tests that allow multiplexing emerged, as antibody seropositivity can instruct about individual immunity after an infection with SARS-CoV-2 or after vaccination. As many commercial antibody tests are either time-consuming or tend to produce false negative or false positive results when only one antigen is considered, we developed an automated, flow-based chemiluminescence microarray immunoassay (CL-MIA) that allows for the detection of IgG antibodies to SARS-CoV-2 receptor-binding domain (RBD), spike protein (S1 fragment), and nucleocapsid protein (N) in human serum and plasma in less than 8 min. The CoVRapid CL-MIA was tested with a set of 65 SARS-CoV-2 serology positive or negative samples, resulting in 100% diagnostic specificity and 100% diagnostic sensitivity, thus even outcompeting commercial tests run on the same sample set. Additionally, the prospect of future quantitative assessments (i.e., quantifying the level of antibodies) was demonstrated. Due to the fully automated process, the test can easily be operated in hospitals, medical practices, or vaccination centers, offering a valuable tool for COVID-19 serosurveillance.

Ultrasensitive and Simultaneous Detection of Two Specific SARS-CoV-2 Antigens in Human Specimens Using Direct/Enrichment Dual-Mode Fluorescence Lateral Flow Immunoassay

“…Many SARS-CoV-2 variants (e.g., B.1.1.7, B.1.351, and P.1) have rapidly emerged around the world, with several mutations in the functional domain of S protein as well as in other proteins (including NP). , These variants raise important concerns about their immune evasion potential and the risk of false-negative results, which pose a growing challenge for the accurate and sensitive detection of SARS-CoV-2 via antigens. Some research studies have demonstrated that the combination of multiple target detection has outstanding performance (improving the sensitivity or accuracy) in SARS-CoV-2 and its variant diagnosis. − In theory, the simultaneous detection of SARS-CoV-2 S and NP antigens offers tremendous potential for improving the accuracy and detection rate of SARS-CoV-2 infection in the point-of-care testing (POCT) area.…”

Multiplexed flow cytometric approach for detection of anti-SARS-CoV-2 IgG, IgM and IgA using beads covalently coupled to the nucleocapsid protein

“…Other multiplex approaches target the detection of IgG antibody against SARS‐CoV‐2 antigens were also described, such as a flow‐based chemiluminescence microarray immunoassay (CL‐MIA) (Klüpfel et al . 2021 ) and the VaxArray Coronavirus SeroAssay Kit (Dawson et al . 2021 ).…”

Design and Implementation of a Miniaturized Dual-wavelength UV Crosslinker

“…In the past few decades, the biomolecule microarray has emerged as a high-throughput and parallel screening tool for different types of optical sensing techniques identifying biomolecular interactions, including surface plasmon resonance imaging (SPRi), fluorescence imaging, surface-enhanced Raman scattering (SERS), and chemiluminescence. (1)(2)(3)(4) During the identification, different reagents injected during the interactions can produce varied signal changes in spots of the microarray and their background. To maximize the contrast between the signals from the spots and their background, physical crosslinking immobilization has been widely used in microarray fabrication owing to its ability to conjugate a large amount of a reagent to a substrate in a short time.…”

“…To maximize the contrast between the signals from the spots and their background, physical crosslinking immobilization has been widely used in microarray fabrication owing to its ability to conjugate a large amount of a reagent to a substrate in a short time. (3)(4)(5)(6) Currently, the most used crosslinking techniques can be categorized as chemical and physical methods. In the chemical crosslinking methods, polycondensation and polyaddition are the most common reactions for the production of polymers.…”