Development of in-house ELISAs for the detection of anti-SARSCoV2 RBD and N IgG and IgM antibodies in biological samples.
J King Saud Univ Sci
; 33(4): 101439, 2021 Jun.
Article
in English
| MEDLINE | ID: covidwho-1185114
ABSTRACT
By the end of year 2019, the new virus SARS-CoV-2 appeared, causing the Coronavirus Disease 2019 (COVID-19), and spread very fast globally. A continuing need for diagnostic tools is a must to contain its spread. Till now, the gold standard method, the reverse transcription polymerase chain reaction (RT-PCR), is the precise procedure to detect the virus. However, SARS-CoV-2 may escape RT-PCR detection for several reasons. The development of well-designed, specific and sensitive serological test like enzyme immunoassay (EIA) is needed. This EIA can stand alone or work side by side with RT-PCR. In this study, we developed several EIAs including plates that are coated with either specially designed SARS-CoV-2 nucleocapsid or surface recombinant proteins. Each protein type can separately detect anti-SARS-CoV-2 IgM or IgG antibodies. For each EIAs, the cut-off value, specificity and sensitivity were determined utilizing RT-PCR confirmed Covid-19 and pre-pandemic healthy and other viruses-infected sera. Also, the receiver operator characteristic (ROC) analysis was performed to define the specificities and sensitivities of the optimized assay. The in-house EIAs were validated by comparing against commercial EIA kits. All in-house EIAs showed high specificity (98-99%) and sensitivity (97.8-98.9%) for the detection of IgG/IgM against RBD and N proteins of SARS-CoV-2. From these results, the developed Anti-RBD and anti-N IgG and IgM antibodies EIAs can be used as a specific and sensitive tool to detect SARS-CoV-2 infection, calculate the burden of disease and case fatality rates.
AUC, Area under the curve; BSA, Bovine serum albumin; CMV, Cytomegalovirus; CoVs, Coronaviruses; Covid-19; Covid-19, Coronavirus Disease-2019; E, Envelope protein; EIA, Enzyme immunoassay; ELISA; ELISA, Enzyme-linked immunosorbant assay; HBV, Hepatitis B virus; HCV, Hepatitis C virus; HIV, Human immunodeficiency virus; HTLV, Human T-lymphotropic virus; M, Membrane protein; N, Nucleocapsid protein; Nucleocapsid protein; OD, Optical density; ORFs, Open reading frames; RBD, Receptor-binding domain; ROC, Receiver operating characteristic; RT-PCR, Reverse transcription polymerase chain reaction; S, Spike protein; SARS-CoV-2; SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2; SD, Standard deviations; Spike protein; TMB, 3,3',5,5'-Tetramethylbenzidine
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Type of study:
Diagnostic study
/
Prognostic study
Language:
English
Journal:
J King Saud Univ Sci
Year:
2021
Document Type:
Article
Affiliation country:
J.jksus.2021.101439
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