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Clin Chem . Evaluation of Orthogonal Testing Algorithm for Detection of SARS-CoV-2 IgG Antibodies

tetano

Editor, Senior Moderator
Clin Chem


. 2020 Sep 7;hvaa210.
doi: 10.1093/clinchem/hvaa210. Online ahead of print.
Evaluation of Orthogonal Testing Algorithm for Detection of SARS-CoV-2 IgG Antibodies


Gang Xu[SUP] 1 [/SUP], Anthony J Emanuel[SUP] 1 [/SUP], Satish Nadig[SUP] 2 [/SUP], Shikhar Mehrotra[SUP] 2 [/SUP], Brittany A Caddell[SUP] 1 [/SUP], Scott R Curry[SUP] 3 [/SUP], Frederick S Nolte[SUP] 1 [/SUP], Nikolina Babic[SUP] 1 [/SUP]



Affiliations

Abstract

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody testing is an important tool in assessment of pandemic progress, contact tracing, and identification of recovered coronavirus disease 2019 (COVID-19) patients. We evaluated an orthogonal testing algorithm (OTA) to improve test specificity in these use cases.
Methods: A two-step OTA was applied where individuals who initially tested positive were tested with a second test. The first-line test, detecting IgG antibodies to the viral nucleocapsid protein was validated in 130 samples and the second-line test, detecting IgG antibodies to the viral spike protein in 148 samples. The OTA was evaluated in 4,333 clinical patient specimens. The seropositivity rates relative to the SARS-CoV-2 PCR positivity rates were evaluated from our entire patient population data (n = 5,102).
Results: The first-line test resulted in a clinical sensitivity of 96.4% (95% CI; 82.3% to 99.4%), and specificity of 99.0% (95% CI; 94.7% to 99.8%), whereas the second-line test had a sensitivity of 100% (95% CI; 87.7% to 100%) and specificity of 98.4% (95% CI; 94.2% to 99.5%). Using the OTA, 78/98 (80%) of initially positive SARS-CoV-2 IgG results were confirmed with a second-line test, while 11/42 (26%) of previously diagnosed COVID-19 patients had no detectable antibodies as long as 94 days post PCR diagnosis.
Conclusion: Our results show that an OTA can be used to identify patients who require further follow-up due to potential SARS CoV-2 IgG false positive results. In addition, serological testing may not be sufficiently sensitive to reliably detect prior COVID-19 infection.
 
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