A multiplex antigen microarray for simultaneous IgG and IgM detection against SARS‐CoV‐2 reveals higher seroprevalence than reported

Summary The surge of SARS‐CoV‐2 has challenged health systems worldwide and efficient tests to detect viral particles, as well as antibodies generated against them, are needed. Specificity, sensitivity, promptness or scalability are the main parameters to estimate the final performance, but rarely all of them match in a single test. We have developed SCOVAM, a protein microarray with several viral antigens (spike, nucleocapsid, main protease Nsp5) as capturing probes in a fluorescence immunoassay for COVID‐19 serological testing. SCOVAM depicts IgG and IgM antibody responses against each of these proteins of 22 individuals in a single microscope slide. It detects specific IgM (0.094 μg ml‐1) and IgG (~0.017 μg ml‐1) and is scalable and cost‐effective. We validated SCOVAM by comparing with a widely used chemiluminescent commercial serological test (n = 742). SCOVAM showed twice the sensitivity and allowed following seroconversion in a single assay. By analysing the prevalence 4 months later in a subset of 76 positive sera, we still detected 93.42% of positives, almost doubling the detection of the commercial assay. The higher sensitivity of SCOVAM is especially relevant to screen sera for convalescent plasma‐based treatments, high‐throughput antibody response monitoring after vaccination or evaluation of vaccine efficiency. We have developed a test to discriminate antibodies against SARS‐CoV‐2 in the blood serum. It detects IgM and IgG antibodies against the Spike, Nucleocapsid and Main protease of the virus, simultaneously. We printed these viral proteins on slides forming microarrays and incubated them with human sera for fluorescent detection. We established the limit of detection and an algorithm for automated diagnostics. Our test resulted twice as sensitive as a commercial test, being able to better follow antibody response through time, what will be very useful to track the epidemiology of COVID‐19 and the effectivity of the developed vaccines.