scholarly journals Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS‐CoV‐2 Neutralizing Antibodies

2020 ◽  
Vol 131 (1) ◽  
Author(s):  
Abigail Vanderheiden ◽  
Venkata Viswanadh Edara ◽  
Katharine Floyd ◽  
Robert C. Kauffman ◽  
Grace Mantus ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Focus Reduction

Development of a focus reduction neutralization test (FRNT) for detection of mumps virus neutralizing antibodies

2010 ◽  
Vol 163 (1) ◽  
pp. 153-156 ◽  
Author(s):  
Sunil R. Vaidya ◽  
David W.G. Brown ◽  
Li Jin ◽  
Dhanraj Samuel ◽  
Nick Andrews ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Mumps Virus ◽  
Focus Reduction

scholarly journals Presence and short-term persistence of SARS-CoV-2 neutralizing antibodies in COVID-19 convalescent plasma donors.

2020 ◽  
Author(s):  
Kyle Annen ◽  
Thomas E Morrison ◽  
Melkon G DomBourian ◽  
Mary K McCarthy ◽  
Leah Huey ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Neutralizing Antibody ◽  
Considerable Variability ◽  
Binding Assays ◽  
Short Term ◽  
Live Virus ◽  
Donor Population ◽  
Antibody Titers ◽  
Focus Reduction

In March 2020, the FDA approved the use of COVID-19 convalescent plasma (CCP) as an investigational new drug for treatment of COVID-19. Since then, collection of CCP from COVID-19 recovered patients has been implemented in several donor centers across the country. Childrens Hospital Colorado rapidly put into practice a CCP collection protocol, necessitating the development and implementation of assays to evaluate SARS-CoV-2 antibodies in CCP units. We evaluated 87 separate units of CCP collected from 36 donors over two to four sequential donations using both antigen-binding assays for SARS-CoV-2 nucleoprotein and spike antigens, and a live virus focus reduction neutralization test (FRNT50). Our data shows that the majority of donors (83 percent) had a FRNT50 titer of 1/80 or greater, and 61 percent had a titer greater than or equal to 1/160, which meet the FDA criteria for acceptable CCP units. Additionally, our data indicates that analysis of antibodies to a single SARS-CoV-2 antigen is likely to miss a percentage of seroconverters. These individuals, however, tend to have neutralizing antibody titers of less than 1/80. Of note, there was considerable variability in the short term, sustained antibody response, measured by neutralizing antibody titers, among our donor population.

scholarly journals Five Commercial Immunoassays for SARS-CoV-2 Antibody Determination and Their Comparison and Correlation with the Virus Neutralization Test

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 593
Author(s):  
Václav Šimánek ◽  
Ladislav Pecen ◽  
Zuzana Krátká ◽  
Tomáš Fürst ◽  
Hana Řezáčková ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Young Patients ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Age Dependent ◽  
Previous Contact ◽  
Dependent Elderly ◽  
Protein Assays ◽  
Antibody Determination

There is an ongoing debate as to whether SARS-CoV-2 antibodies can be found in patients who have recovered from COVID-19 disease. Currently, there is no consensus on whether the antibodies, if present, are protective. Our regular measurements of SARS-CoV-2 antibodies, starting in July 2020, have provided us with the opportunity of becoming acquainted with the five different immunoassays. A total of 149 patients were enrolled in our study. We measured the samples using each immunoassay, then performing a virus neutralization test and comparing the results of SARS-CoV-2 antibodies with this test. We observed that the production of neutralizing antibodies is age-dependent. Elderly patients have a higher proportion of high neutralizing titers than young patients. Based on our results, and in combination with the literature findings, we can conclude that the serological SARS-CoV-2 antibody measurement is a helpful tool in the fight against COVID-19. The assays can provide information about the patient’s previous contact with the virus. Anti-spike protein assays correlate well with the virus neutralization test and can be used in the screening of potential convalescent plasma donors.

scholarly journals Development of SARS-CoV-2 Specific IgG and Virus-Neutralizing Antibodies after Infection with Variants of Concern or Vaccination

Vaccines ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 700
Author(s):  
Franziska Neumann ◽  
Ruben Rose ◽  
Janine Römpke ◽  
Olaf Grobe ◽  
Thomas Lorentz ◽  
...  
Keyword(s):  
Immune Response ◽  
Receptor Binding ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Vaccine Dose ◽  
High Avidity ◽  
Receptor Binding Domain ◽  
Robust Immune Response ◽  
Specific Igg ◽  
Nucleoprotein N

The humoral immunity after SARS-CoV-2 infection or vaccination was examined. Convalescent sera after infection with variants of concern (VOCs: B.1.1.7, n = 10; B.1.351, n = 1) and sera from 100 vaccinees (Pfizer/BioNTech, BNT162b2, n = 33; Moderna, mRNA-1273, n = 11; AstraZeneca, ChAdOx1 nCoV-19/AZD1222, n = 56) were tested for the presence of immunoglobulin G (IgG) directed against the viral spike (S)-protein, its receptor-binding domain (RBD), the nucleoprotein (N) and for virus-neutralizing antibodies (VNA). For the latter, surrogate assays (sVNT) and a Vero-cell based neutralization test (cVNT) were used. Maturity of IgG was determined by measuring the avidity in an immunoblot (IB). Past VOC infection resulted in a broad reactivity of anti-S IgG (100%), anti-RBD IgG (100%), and anti-N IgG (91%), while latter were absent in 99% of vaccinees. Starting approximately two weeks after the first vaccine dose, anti-S IgG (75–100%) and particularly anti-RBD IgG (98–100%) were detectable. After the second dose, their titers increased and were higher than in the convalescents. The sVNT showed evidence of VNA in 91% of convalescents and in 80–100%/100% after first/second vaccine dose, respectively. After the second dose, an increase in VNA titer and IgGs of high avidity were demonstrated by cVNT and IB, respectively. Re-vaccination contributes to a more robust immune response.

scholarly journals Seroprevalence and correlates of SARS-CoV-2 neutralizing antibodies from a population-based study in Bonn, Germany

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
N. Ahmad Aziz ◽  
Victor M. Corman ◽  
Antje K. C. Echterhoff ◽  
Marcel A. Müller ◽  
Anja Richter ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Response Rate ◽  
Neutralization Test ◽  
Population Based ◽  
Immunofluorescence Assay ◽  
Test Results ◽  
Community Based ◽  
Population Based Study ◽  
Temporal Course ◽  
Confirmatory Tests

AbstractTo estimate the seroprevalence and temporal course of SARS-CoV-2 neutralizing antibodies, we embedded a multi-tiered seroprevalence survey within an ongoing community-based cohort study in Bonn, Germany. We first assessed anti-SARS-CoV-2 immunoglobulin G levels with an immunoassay, followed by confirmatory testing of borderline and positive test results with a recombinant spike-based immunofluorescence assay and a plaque reduction neutralization test (PRNT). Those with a borderline or positive immunoassay result were retested after 4 to 5 months. At baseline, 4771 persons participated (88% response rate). Between April 24th and June 30th, 2020, seroprevalence was 0.97% (95% CI: 0.72−1.30) by immunoassay and 0.36% (95% CI: 0.21−0.61) when considering only those with two additional positive confirmatory tests. Importantly, about 20% of PRNT+ individuals lost their neutralizing antibodies within five months. Here, we show that neutralizing antibodies are detectable in only one third of those with a positive immunoassay result, and wane relatively quickly.

scholarly journals A GFP Reporter MR766-Based Flow Cytometry Neutralization Test for Rapid Detection of Zika Virus-Neutralizing Antibodies in Serum Specimens

Vaccines ◽  
2019 ◽  
Vol 7 (3) ◽  
pp. 66 ◽  
Author(s):  
Frumence ◽  
Viranaicken ◽  
Gadea ◽  
Desprès
Keyword(s):  
Flow Cytometry ◽  
Zika Virus ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Health Concern ◽  
Structural Protein ◽  
Adult Mice ◽  
Gfp Reporter ◽  
Infected Cells ◽  
High Level

Zika virus (ZIKV) is an emerging arthropod-borne virus of major public health concern. ZIKV infection is responsible for congenital Zika disease and other neurological defects. Antibody-mediated virus neutralization is an essential component of protective antiviral immunity against ZIKV. In the present study, we assessed whether our GFP reporter ZIKV derived from African viral strain MR766 could be useful for the development of a flow cytometry neutralization test (FNT), as an alternative to the conventional plaque-reduction neutralization test (PRNT). To improve the efficacy of GFP-expressing MR766, we selected virus variant MR766GFP showing a high level of GFP signal in infected cells. A MR766GFP-based FNT was assayed with immune sera from adult mice that received ZIKBeHMR-2. The chimeric ZIKV clone ZIKBeHMR-2 comprises the structural protein region of epidemic strain BeH819015 into MR766 backbone. We reported that adult mice inoculated with ZIKBeHMR-2 developed high levels of neutralizing anti-ZIKV antibodies. Comparative analysis between MR766GFP-based FNT and conventional PRNT was performed using mouse anti-ZIKBeHMR-2 immune sera. Indistinguishable neutralization patterns were observed when compared with PRNT50 and FNT50. We consider that the newly developed MR766GFP-based FNT is a valid format for measuring ZIKV-neutralizing antibodies in serum specimens.

scholarly journals Evidence of neutralizing antibodies against SARS-CoV-2 in domestic cats living with owners with a history of COVID-19 in Lima, Peru

2021 ◽  
Author(s):  
Luis M. Jara ◽  
Cusi Ferradas ◽  
Francesca Schiaffino ◽  
Camila Sanchez-Carrion ◽  
Ana Martinez ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Domestic Animals ◽  
Disease Prevalence ◽  
Receptor Binding Domain ◽  
Domestic Cats ◽  
Viral Reservoirs ◽  
History Of ◽  
Community Transmission ◽  
Disease Exposure

SARS-CoV-2 can infect a variety of wild and domestic animals worldwide. Of these, domestic cats are highly susceptible species and potential viral reservoirs. As such, it is important to investigate disease exposure in areas with active community transmission and high disease prevalence. In this report we demonstrate the presence of serum neutralizing antibodies against the receptor binding-domain (RBD) of the SARS-CoV-2 in cats whose owners had been infected with SARS-CoV-2 in Lima, Peru, using a commercial competitive ELISA SARS-CoV-2 Surrogate Virus Neutralization Test. Out of 41 samples, 17.1% (7/41) and 31.7% (13/41) were positive, using the cut-off inhibition value of 30% and 20%, respectively. Not all cats living in a single house had detectable neutralizing antibodies showing that heterogenous exposure and immune among cohabiting animals. This is the first report of SARS-COV-2 exposure of domestic cats in Lima, Peru. Further studies are required to ascertain the prevalence of SARS-COV-2 exposure among domestic cats of Lima, Peru.

scholarly journals Molecular and seroepidemiologic studies of Enterovirus 71 infection in the State of Pará, Brazil

2005 ◽  
Vol 47 (2) ◽  
pp. 65-71 ◽  
Author(s):  
Ceyla M.O. Castro ◽  
Ana Cecília R. Cruz ◽  
Edson E. da Silva ◽  
Maria de Lourdes C. Gomes
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Neurological Diseases ◽  
Enterovirus 71 ◽  
Foot And Mouth Disease ◽  
Northern Region ◽  
Molecular Pattern ◽  
Seroepidemiologic Studies ◽  
Stool Samples ◽  
The City

In many countries, the Enterovirus 71 (EV-71) Picornaviridae family is associated to hand, foot and mouth disease in addition to acute neurological diseases while in Brazil these viruses are more closely associated to the latter group. The aim of this research was to use the first EV-71 isolate of the Northern region of Brazil in molecular and seroepidemiologic studies. Two (2.2%) out of 88 stool samples (44 cases of AFP), collected from January 1998 to December 2000 were positive for EV-71 isolation (73442/PA/99). Nucleotide sequence of the gen that codifies the VP1 protein showed that isolate 73442/PA/99 was similar to the EV-71 strains belonging to genotype B - more closely identified with EV-71 from North America. Neutralization test with 389 sera samples collected from January 1998 to November 2001, from individuals ranging from 0 to 15 years of age living in the city of Belém, State of Pará showed the following results in relation to isolate 73442/PA/99 and prototype BrCr: a total of 207 individuals (53.2%) had neutralization antibodies to both viruses, 167 (42.9%) had no antibodies and 15 showed the presence of neutralizing antibodies to one of the two viruses. Only 20.2% of the children aged 0 to 3 had neutralizing antibodies to EV-71, indicating that these children were more susceptible to the infection. Both the seroprevalence study and VP1 sequencing were important to demonstrate the spread and the molecular pattern of the EV-71 circulating in the Northern Region of Brazil.

scholarly journals Development of specific immunity in laboratory animals after co-immunization against seasonal influenza and COVID-19

2022 ◽  
Vol 98 (6) ◽  
pp. 648-656
Author(s):  
G. M. Ignatyev ◽  
I. A. Leneva ◽  
A. V. Atrasheuskaya ◽  
L. I. Kozlovskaya ◽  
N. P. Kartashova ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Seasonal Influenza ◽  
Neutralization Test ◽  
Elisa Test ◽  
Control Measures ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Post Vaccination ◽  
Specific Immunity ◽  
Influenza Prevention

Introduction. In clinical practice, the differential diagnosis of COVID-19 can be challenging during the flu season, entailing serious consequences such as delays in appropriate control measures against the SARS-CoV-2 pandemic. Another problem is posed by co-infection of SARS-CoV-2 and influenza virus (IV), which significantly contributes to the severity of the COVID-19 disease. This study was aimed to explore the cross-impact of co-administration of Russian influenza and COVID-19 vaccines on development of specific immunity in laboratory animals.Materials and methods. The study was conducted on BALB/c mice. The animals were inoculated intramuscularly with the vaccine for COVID-19 prevention (CoviVac) and the vaccine for influenza prevention (Flu-M). The sera from the immunized animals were examined separately. Three IV strains were used in the hemagglutination inhibition assay. Antibodies (Abs) against SARS-CoV-2 were detected by an enzyme-linked immunosorbent assay (ELISA). The neutralization test was performed to detect virus neutralizing antibodies against SARS-CoV-2 and IV.Results. Relatively high titers of specific Abs were found in the groups of animals inoculated with one vaccine and with two vaccines concurrently. In the groups of animals inoculated with CoviVac and with two vaccines concurrently, both in the ELISA test and in the neutralization test, the average titers of specific Abs against SARSCoV- 2 did not demonstrate any statistical difference. The group of animals inoculated concurrently with two vaccines demonstrated statistically higher titers of Abs against IV after the second immunization compared to the group of animals inoculated with Flu-M.Discussion. The study has shown that post-vaccination immunity both to IV and to SARS-CoV-2 develops after co-vaccination with two vaccines. The observed enhanced post-vaccination immune response to IV in the coimmunized laboratory animals needs further research.Conclusion. The performed studies suggest the possibility of co-administration of two vaccines to prevent influenza and COVID-19.

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