Development and validation of an enzyme-linked immunoassay kit for diagnosis and surveillance of COVID-19

Bagno, Flávia Fonseca; Sérgio, Sarah A. R.; Figueiredo, María Marta; Godói, Lara Carvalho; Andrade, Luis Adan Flores; Salazar, Natália C.; Soares, Camila Pereira; Aguiar, Andressa; Almeida, Flávia Jaqueline; Silva, Edimilson D. da; Ferreira, Antônio G. P.; Durigon, Edison Luiz; Gazzinelli, Ricardo T.; Teixeira, Santuza Maria Ribeiro; Fernandes, Ana Paula; Fonseca, Flávio Guimarães da

doi:10.1016/j.jcvp.2022.100101

Cited by 8 publications

(3 citation statements)

References 28 publications

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“…Serum samples were initially tested for the detection of IgG anti-SARS-CoV-2 antibodies using an enzyme-linked immunosorbent assay (ELISA) developed by the Vaccine Technology Centre of the Federal University of Minas Gerais [22]. The test detected antibodies against the Nucleocapsid (N) protein of SARS-CoV-2.…”

Section: Enzyme Immunoassay (Elisa)mentioning

confidence: 99%

Seroprevalence of SARS-CoV-2 in hospital workers in the southern region of Minas Gerais state in Brazil: An analysis of the pre-vaccine period

et al. 2023

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The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection seroprevalence can be performed by detecting anti-SARS-CoV-2 antibodies. The survey is essential to understand the disease transmission's dynamic in the studied population. This study aimed to carry out a seroepidemiological survey of SARS-CoV-2 in three hospitals located in the south of Minas Gerais state, Brazil. 859 samples were collected from August to December 2020 when SARS-CoV-2 vaccines were still not available and Enzyme-linked immunosorbent assays (ELISA) were performed on participants sera. The average age of participants was 38 years, and most were women (71.4%). Likewise, most participants were classified as health professionals with direct or indirect contact with patients with COVID-19 (74.5%). The other participants tested belonged to other sectors, such as the administrative one (11,6%). Considering clinical symptoms, 15.8% of participants reported diarrhoea, 6.4% fever, 5.8% respiratory distress, and 7.0% loss of smell and taste. Many participants reported contact with infected patients (63.35%). Regarding the ELISA tests, 21.6% of the participants had positive results and hospital 3 had the highest positivity (21.7%), followed by hospital 2 (21.6%) and hospital 1 (20.3%). The prevalence was higher in women compared to men (22,8% and 18,7%, respectively). Regarding the area of expertise, the highest positivity (20.9%) was observed among health professionals. However, professionals who worked exclusively with COVID-19 had lower positivity when compared to professionals who did not work directly with respectively). When analysing the correlation between the ELISA tests with the other variables, a significant association was detected with these previous serological variables, previous contact with COVID-19 and the presence of fever symptoms, loss of smell and taste. Clinical symptoms associated with serological tests are important tools for monitoring the disease among health professionals.

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Section: Enzyme Immunoassay (Elisa)mentioning

confidence: 99%

Seroprevalence of SARS-CoV-2 in hospital workers in the southern region of Minas Gerais state in Brazil: An analysis of the pre-vaccine period

et al. 2023

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“…All three proteins were expressed in Escherichia coli strain BL21(DE3). The recombinant antigens were purified with affinity chromatography using nickel columns in an AKTA Prime plus system following the manufacturer's instructions (GE Healthcare, Chicago, IL, USA) [21]. The eluted fractions containing the purified proteins were analyzed using SDS-PAGE, Western blot with Anti-His Tag antibody (Supplementary Figure S1).…”

Section: Rt-qpcr and Rna Sequencingmentioning

confidence: 99%

SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation

Coelho,

da Silva,

Lopes

et al. 2023

Microorganisms

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SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants—Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients’ follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.

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“…Humoral immune responses to the SARS-CoV2 S protein are typically evaluated by Enzyme Linked Immunosorbent Assay (ELISA) and its many variants (CLIA, LFA, etc.) (Bagno et al, 2022). However, conventional laboratory testing on plasma or serum samples is not always readily available.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of dried blood spot-measured anti-SARS-CoV2 Spike IgG in mRNA-vaccinated healthcare workers

Puccini

Fantini²,

Biagetti³

et al. 2023

Front. Microbiol.

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IntroductionOne of the major criticisms facing the research community during SARS-CoV2 pandemic was the lack of large-scale, longitudinal data on the efficacy of the SARS-CoV2 mRNA vaccines. Currently, even if COVID-19 antiviral treatments have been authorized by European Medicine Agency, prevention through approved specific vaccines is the best approach available in order to contain the ongoing pandemic.ObjectivesHere, we studied the antibody kinetic over a one-year period from vaccination with the Pfizer-BioNTech (Pfizer) vaccines and subsequent boosting with either the BioNTech or Moderna (Spikevax) vaccines in a large cohort of 8,071 healthcare workers (HCW). We also described the impact of SARS-CoV2 infection on antibody kinetic over the same period.MethodsWe assessed the anti SARS-CoV2 Spike IgG antibody kinetic by the high throughput dried blood spot (DBS) collection method and the GSP®/DELFIA® Anti-SARS-CoV2 IgG assay (PerkinElmer®).ResultsOur data support existing models showing that SARS-CoV2 vaccination elicits strong initial antibodies responses that decline with time but are transitorily increased by administering a vaccine booster. We also showed that using heterologous vaccine/booster combinations a stronger antibody response was elicited than utilizing a booster from the same vaccine manufacturer. Furthermore, by considering the impact of SARS-CoV2 infection occurrence in proximity to the scheduled booster administration, we confirmed that booster dose did not contribute significantly to elicit higher antibody responses.ConclusionDBS sampling in our large population of HCWs was fundamental to collect a large number of specimens and to clarify the effective mRNA vaccine-induced antibody kinetic and the role of both heterologous boosters and SARS-CoV2 infection in modulating antibody responses.

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Development and validation of an enzyme-linked immunoassay kit for diagnosis and surveillance of COVID-19

Cited by 8 publications

References 28 publications

Seroprevalence of SARS-CoV-2 in hospital workers in the southern region of Minas Gerais state in Brazil: An analysis of the pre-vaccine period

Seroprevalence of SARS-CoV-2 in hospital workers in the southern region of Minas Gerais state in Brazil: An analysis of the pre-vaccine period

SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation

Kinetics of dried blood spot-measured anti-SARS-CoV2 Spike IgG in mRNA-vaccinated healthcare workers

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