Establishment and performance evaluation of an immunoassay for the antibody detection against the receptor binding domain of the SARS-CoV-2 spike protein
DOI:
https://doi.org/10.51481/amc.v65i4.1355Keywords:
SARS CoV-2, COVID-19, antibodies, inmunoassay, enzyme-linked immunosorbent assayAbstract
Aim: To establish a semiquantitative immunoassay for antibody detection against the RBD of the severe acute respiratory syndrome coronavirus 2 spike protein and to evaluate its performance to be used as a diagnostic supporting tool.
Methods: A recombinant severe acute respiratory syndrome coronavirus 2 spike protein was produced. This protein was used as antigenic substrate in two semiquantitative enzyme-linked immunoassays for the detection of human immunoglobulins M and immunoglobulins G. A set of serum samples (N=129) from patients with prior viral infection confirmed by reverse transcription polymerase chain reaction, processed between August 2020 and November 2021, were used as positive samples. A panel of pre-pandemic samples (N=196), obtained prior to December 2019, were used as negative samples to evaluate the assay performance. Multiple samples from 99 volunteers were used to examine test response to seroconversion. The interference between seropositivity against severe acute respiratory syndrome coronavirus 2 and dengue virus was also evaluated.
Results: The immunoglobulin G detection assay showed 81.4% sensitivity, 86.2% specificity, and positive and negative predictive values of 79.5% and 87.6% respectively. The immunoglobulin M detection assay yielded 72.1% sensitivity, 54.1% specificity, and positive and negative predictive values of 25.6% and 89.8% respectively. No significant differences were found between the measurements according to sex or linear correlation between this variable and age. The values presented significant differences according to the condition of self-reported presence or absence of COVID-19 like symptoms. No correlation was found between seropositivity for severe acute respiratory syndrome coronavirus 2 and dengue virus.
The immunoglobulin G detection assay generated lower but constant values on samples from voluntary donors who reported not having any contact with the virus compared to samples
from donors exposed to it, and high but variable values in magnitude on samples from vaccinated volunteers or those with previous severe acute respiratory syndrome coronavirus 2 infection compared to samples from donors without exposure to the viral antigen.
Conclusions: Our established immunoglobulin M detection assay presents poor diagnostic value. On the other hand, the immunoglobulin G detection assay shows satisfactory performance, and coheres to the behavior reported for this type of test according to the demographic and clinic characteristics of the volunteer, so it could be used as a reliable and practical tool in clinical applications and as diagnostic complement. It is necessary to develop more studies on cross-reactions of antibodies against severe acute respiratory syndrome coronavirus 2 with other entities of clinical interest and present in our tropical area.
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