The extent of SARS-CoV-2 infection throughout the United States population is currently unknown. High quality serology is key to avoiding medically costly diagnostic errors, as well as to assuring properly informed public health decisions. Here, we present an optimized ELISA-based serology protocol, from antigen production to data analyses, that helps define thresholds for IgG and IgM seropositivity with high specificities. Validation of this protocol is performed using traditionally collected serum as well as dried blood on mail-in blood sampling kits. Archival (pre-2019) samples are used as negative controls, and convalescent, PCR-diagnosed COVID-19 patient samples serve as positive controls. Using this protocol, minimal cross-reactivity is observed for the spike proteins of MERS, SARS1, OC43 and HKU1 viruses, and no cross reactivity is observed with anti-influenza A H1N1 HAI. Our protocol may thus help provide standardized, population-based data on the extent of SARS-CoV-2 seropositivity, immunity and infection. Understanding the infection parameters and host responses against SARS-CoV-2 require data from large cohorts using standardized methods. Here, the authors optimize a serum ELISA protocol that has minimal cross-reactivity and flexible sample collection workflow in an attempt to standardize data generation and help inform on COVID-19 pandemic and immunity.
【저자키워드】 antibodies, viral infection, ELISA, Laboratory techniques and procedures, 【초록키워드】 viruses, public health, SARS-CoV-2, protocol, Immunity, serology, SARS-COV-2 infection, COVID-19 pandemic, Infection, diagnostic, host response, MERS, Antigen, cross-reactivity, serum, validation, Seropositivity, understanding, H1N1, large cohort, United States, convalescent, threshold, OC43, HKU1, Blood, Cross reactivity, SARS1, Spike proteins, COVID-19 patient, host responses, IgG and IgM, negative controls, help, Specificities, thresholds, parameter, positive controls, flexible, performed, collected, the United State, the spike protein, costly, analyses, ELISA protocol, HAI, 【제목키워드】 pandemic, Serosurvey, Blood, ELISA protocol, the SARS-CoV-2,