Comparative Analysis of the Analytical Sensitivity of ELISA Test System DIA®-SARS-CoV-2-Ag-R with Rapid Tests for Viral Antigen SARS-CoV-2 Detection
Authors:   PJSC SPC “Diaproph-Med“, Horlov А.Y. , Serdiuk V.H. ,   PJSC SPC “Diaproph-Med“, Kiselova О.K. ,   PJSC SPC “Diaproph-Med“, Shevchuk A.O. ,   PJSC SPC “Diaproph-Med“, Melnykova O.I. ,   PJSC SPC “Diaproph-Med“, Chumak O.M. ,   PJSC SPC “Diaproph-Med“, Horlov Y.I. ,   PJSC SPC “Diaproph-Med“, Shevchuk O.A. ,   PJSC SPC “Diaproph-Med“, Spivak M.Ya. ,   Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
Published: June 2021

A novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease (COVID)-19, that emerged as a major pandemic. SARS-CoV-2 was identified as a betacoronavirus. Nucleocapsid protein (NP) is evolutionary high protein homologies and solid structure protein for SARS-CoV-2 detection as opposed to other proteins, that aren`t reliable as a single viral antigen during diagnostics methods. The viral RNA can be detected from nasal and pharyngeal swabs and bronchoalveolar lavage samples by PCR assay. However, the wrong collection of samples can lead to false-negative diagnosis and have dangerous consequences at this stage of pandemic. One of efficient and accurate methodological approaches for the screening of pathogens are serological assays. Aim. Evaluation and comparison of the sensitivity of invented DIA®-SARS-CoV-2-Ag-R enzyme-linked immunosorbent assay (ELISA)-based test system and commercial rapid tests, which detect the viral antigen of SARS-CoV-2. Methods. We carried out a comparison of DIA®-SARS-CoV-2-Ag-R and existed commercial test systems, which are used to detect the antigen of SARS-CoV-2 virus. Rapid tests are intended for nasopharyngeal swabs, but we proposed a protein of our own manufacture – recombinant NP as a calibrator. The detection limit was calibrated by standard CFAR #100982 NIBSC, UK. We had determined levels of NP (1400, 900, 750, 640 and 280 pg/mL) that we used as a sample for the rapid tests. The COVID-19 Ag Rapid Tests were performed according to the manufactures instructions at room temperature. Results. DIA®-SARS-CoV-2-Ag-R detected 10 pg/mL of in-house standard of recombinant SARS-CoV-2 NP. The positive results were observed using 1400, 900, 750 pg/mL, while 640 and 280 pg/mL samples were performed as negative in ABBOTT-PanBio test. The rapid tests manufactured by МBU, BIOTIME, Core Technology, SD BIOSENSOR and Turklab showed positive results only in 1400 pg/mL concentration. NP of lower levels was detected as a negative sample. The LEPU MEDICAL test was evaluated as positive sample using 900 pg/mL. The rapid test manufactured by Green Cross Medical Science Corp. showed negative results for all levels of NP. It can mean that the sensitivity of test is lower and demands higher level of antigen to detect the presence of SARS-CoV-2. Conclusions. The study presented an excellent analytical sensitivity of DIA®-SARS-CoV-2-Ag-R against commercial Antigen rapid tests. Thus, invented ELISA test system can be recommended for widespread usage for detection and confirmation of acute stage of SARS-CoV-2 infection.