A modular microarray imaging system for highly specific COVID-19 antibody testing

Hedde, Per Niklas; Abram, Timothy J.; Jain, Aarti; Nakajima, Rie; Assis, Rafael Ramiro de; Pearce, Trevor; Jasinskas, Algis; Toosky, Melody N; Khan, Saahir; Felgner, Philip L.; Gratton, Enrico; Zhao, Weian

doi:10.1039/d0lc00547a

Cited by 36 publications

(37 citation statements)

References 29 publications

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“…The protein microarray COVAM analysis is a high-throughput serological test for SARS-CoV-2 Ab cross-reactivity with a panel of 61 antigens from 23 strains of 10 respiratory tract infection-causing viruses (22). In this assay, each antigen was printed onto microarrays, probed with human plasma, and analyzed with an ArrayCam imager.…”

Section: Resultsmentioning

confidence: 99%

“…COVAM. Serum coronavirus antigen microarray (COVAM) included 67 antigens across respiratory virus subtypes including 11 antigens from SARS-CoV-2 expressed in either baculovirus or HEK-293 cells as previously detailed (22). These antigens were provided by Sino Biological U.S. Inc. as either catalog products or custom synthesis service products.…”

Section: Methodsmentioning

confidence: 99%

“…Here, we sought to characterize epitopes from SARS-CoV-2 and their correlations with disease severity. Enzyme-linked immunosorbent assays (ELISAs) with phage-displayed epitopes (phage ELISAs) and coronavirus antigen microarray (COVAM) analysis (22) examined plasma samples from COVID-19 patients (n = 86). The results demonstrate that Abs to a specific epitope from N protein plus disease risk factors strongly correlate with COVID-19 disease severity.…”

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confidence: 99%

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Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score

Sen

Sanders

Gabriel

et al. 2021

mSphere

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score

Sen

Sanders

Gabriel

et al. 2021

mSphere

Self Cite

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“… 13 , 14 , 16 , 17 This strategy has been used to differentiate SARS-CoV-2 antibodies from that of other pathogens. 18 However, the sensitivities in these studies are limited perhaps due to the lack of quality assessment for the antigen microarray, due to the decision making based on a single type of immunoglobin, or due to the cross-reactivity to form other common coronaviruses. Thus, this study developed a human coronavirus (HCoV) protein microarray, which includes all of the human coronaviruses to significantly reduce the false-positive results that is quality controlled by immunostaining and receptor bindings.…”

Section: Introductionmentioning

confidence: 99%

Development and Application of Human Coronavirus Protein Microarray for Specificity Analysis

Chou

Santos

et al. 2021

Anal. Chem.

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Coronavirus is an enveloped RNA virus that causes mild to severe respiratory diseases in humans, including HKU1-CoV, 229E-CoV, NL63-CoV, OC43-CoV, SARS-CoV, MERS-CoV, and SARS-CoV-2. Due to the outbreak of SARS-CoV-2, it is important to identify the patients and investigate their immune responses. Protein microarray is one of the best platforms to profile the antibodies in the blood because of its fast, multiplexed, and sensitive nature. To fully understand the immune responses and biological specificities, this study developed a human coronavirus (HCoV) protein microarray and included all seven human coronaviruses and three influenza viruses. Each protein was printed in triplicate and formed 14 identical blocks per array. The HCoV protein microarray showed high reproducibility and sensitivity to the monoclonal antibodies against spike and nucleocapsid protein with detection limits of 10–200 pg. The HCoV proteins that were immobilized on the array were properly folded and functional by showing interactions with a known human receptor, e.g., ACE2. By profiling the serum IgG and IgA from 32 COVID-19 patients and 36 healthy patients, the HCoV protein microarray demonstrated 97% sensitivity and 97% specificity with two biomarkers. The results also showed the cross-reactivity of IgG and IgA in COVID-19 patients to spike proteins from various coronaviruses, including that from SARS-CoV, HKU1-CoV, and OC43-CoV. Finally, an innate immune protein named surfactant protein D showed broad affinities to spike proteins in all human coronaviruses. Overall, the HCoV protein microarray is multiplexed, sensitive, and specific, which is useful in diagnosis, immune assessment, biological development, and drug screening.

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“…Finally, only antibodies to one single antigen can be detected, meaning that false negative results would be obtained if patients formed antibodies to a different antigen that is not tested for [ 11 ]. This problem is overcome by microarrays that allow for multiplex analysis of various antigens, usually within hours [ 12 ]. Table 1 shows a comparative overview over different commercial SARS-CoV-2 antibody tests with respect to assay principle, assay time, tested antigens, and test performance.…”

Section: Introductionmentioning

confidence: 99%

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)

Klüpfel

Koros

Dehne³

et al. 2021

Anal Bioanal Chem

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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.

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A modular microarray imaging system for highly specific COVID-19 antibody testing

Abstract: The TinyArray imager, a robust inexpensive portable imaging device, can detect antibodies against SARS-CoV-2 based on coronavirus antigen microarrays.

Cited by 36 publications

References 29 publications

Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score

Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score

Development and Application of Human Coronavirus Protein Microarray for Specificity Analysis

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)

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