Comparative performance of multiplex salivary and commercially available serologic assays to detect SARS-CoV-2 IgG and neutralization titers

Heaney, Christopher D.; Pisanic, Nora; Randad, Pranay R.; Kruczynski, Kate; Howard, Tyrone; Zhu, Xianming; Littlefield, Kirsten; Patel, Eshan U.; Shrestha, Ruchee; Laeyendecker, Oliver; Shoham, Shmuel; Sullivan, David; Gebo, Kelly; Hanley, Daniel; Redd, Andrew D.; Quinn, Thomas C.; Casadevall, Arturo; Zenilman, Jonathan M.; Pekosz, Andrew; Bloch, Evan M.; Tobian, Aaron A. R.

Oral fluid (hereafter saliva) offers a non-invasive sampling method for the detection of SARS-CoV-2 antibodies. However, data comparing performance of salivary tests against commercially-available serologic and neutralizing antibody (nAb) assays are lacking. This study compared the performance of a multiplex salivary SARS-CoV-2 IgG assay targeting antibodies to nucleocapsid (N), receptor binding domain (RBD) and spike (S) antigens to three commercially-available SARS-CoV-2 serology enzyme immunoassays (EIAs) (Ortho Vitros, Euroimmun, and BioRad) and nAb. Paired saliva and plasma samples were collected from 101 eligible COVID-19 convalescent plasma (CCP) donors >14 days since PCR+ confirmed diagnosis. Concordance was evaluated using positive (PPA) and negative (NPA) percent agreement, overall percent agreement (PA), and Cohen’s kappa coefficient. The range between salivary and plasma EIAs for SARS-CoV-2-specific N was PPA: 54.4–92.1% and NPA: 69.2–91.7%, for RBD was PPA: 89.9–100% and NPA: 50.0–84.6%, and for S was PPA: 50.6–96.6% and NPA: 50.0–100%. Compared to a plasma nAb assay, the multiplex salivary assay PPA ranged from 62.3% (N) and 98.6% (RBD) and NPA ranged from 18.8% (RBD) to 96.9% (S). Combinations of N, RBD, and S and a summary algorithmic index of all three (N/RBD/S) in saliva produced ranges of PPA: 87.6–98.9% and NPA: 50–91.7% with the three EIAs and ranges of PPA: 88.4–98.6% and NPA: 21.9–34.4% with the nAb assay. A multiplex salivary SARS-CoV-2 IgG assay demonstrated comparable performance to three commercially-available plasma EIAs and a nAb assay, and may be a viable alternative to assist in screening CCP donors and monitoring population-based seroprevalence and vaccine antibody response.