scholarly journals Suitability of Two Rapid Lateral Flow Immunochromatographic Assays for Predicting SARS-CoV-2 Neutralizing Activity of Sera

2020 ◽  
Author(s):  
Arantxa Valdivia ◽  
Ignacio Torres ◽  
Victor Latorre ◽  
Clara Frances-Gomez ◽  
Josep Ferrer ◽  
...  
Keyword(s):  
Predictive Value ◽  
Fluorescent Protein ◽  
Neutralizing Antibody ◽  
Antibody Test ◽  
Neutralization Assay ◽  
Positive Control ◽  
Lateral Flow ◽  
Control Line ◽  
Assay Sensitivity ◽  
Neutralizing Activity

Purpose: Assessment of commercial SARS-CoV-2 immunoassays for their capacity to provide reliable information on sera neutralizing activity is an emerging need. We evaluated the performance of two commercially-available lateral flow immunochromatographic assays (LFIC) (Wondfo SARS-CoV-2 Antibody test and the INNOVITA 2019-nCoV Ab test) in comparison with a SARS-CoV-2 neutralization pseudotyped assay for COVID-19 diagnosis in hospitalized patients, and investigate whether the intensity of the test band in LFIC associates with neutralizing antibody (NtAb) titers. Patients and Methods: Ninety sera were included from 51 patients with moderate to severe COVID-19. A green fluorescent protein (GFP) reporter-based pseudotyped neutralization assay (vesicular stomatitis virus coated with SARS-CoV-2 spike protein) was used. Test line intensity was scored using a 4-level scale (0 to 3+). Results: Overall sensitivity of LFIC assays was 91.1% for the Wondfo SARS-CoV-2 Antibody test, 72.2% for the INNOVITA 2019-nCoV IgG, 85.6% for the INNOVITA 2019-nCoV IgM and 92.2% for the NtAb assay. Sensitivity increased for all assays in sera collected beyond day 14 after symptoms onset (93.9%, 79.6%,93.9% and 93.9%, respectively). Reactivities equal to or more intense than the positive control line (≥2+) in the Wondfo assay had a negative predictive value of 100% and a positive predictive value of 96.4% for high NtAb50 titers (≥1/160). Conclusions: Our findings support the use of LFIC assays evaluated herein, particularly the Wondfo test, for COVID-19 diagnosis. We also find evidence that these rapid immunoassays can be used to predict high SARS-CoV-2-S NtAb50 titers.

scholarly journals Performance Evaluation of the BZ COVID-19 Neutralizing Antibody Test for the Culture-Free and Rapid Detection of SARS-CoV-2 Neutralizing Antibodies

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2193
Author(s):  
Bo Kyeung Jung ◽  
Jung Yoon ◽  
Joon-Yong Bae ◽  
Jeonghun Kim ◽  
Man-Seong Park ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Diagnostic Performance ◽  
Neutralization Test ◽  
Neutralizing Antibody ◽  
Antibody Test ◽  
Lower Sensitivity ◽  
Control Line ◽  
Perfect Agreement ◽  
Strong Negative Correlation ◽  
Percent Inhibition

Rapid and accurate measurement of SARS-CoV-2 neutralizing antibodies (nAbs) can aid in understanding the development of immunity against COVID-19. This study evaluated the diagnostic performance of a rapid SARS-CoV-2 nAb detection test called the BZ COVID-19 nAb test BZ-nAb (BZ-nAb; BioZentech). Using the 90% plaque-reduction neutralization test (PRNT-90) as a reference, 104 serum specimens collected from COVID-19-positive and -negative patients were grouped into 40 PRNT-90-positive and 64 PRNT-90-negative specimens. The performance of the BZ-nAb was compared with that of the cPass surrogate virus neutralization test (cPass sVNT; Genscript). The BZ-nAb showed a sensitivity ranging from 92.5%–95.0% and specificity ranging from 96.9%–100%, whereas cPass sVNT showed a sensitivity of 100% (95% confidence interval (CI) 90.5%–100%) and specificity of 98.4% (95% CI, 91.6%–100%). The dilution factor obtained with PRNT-90 showed a stronger correlation with the percent inhibition of cPass sVNT (r = 0.8660, p < 0.001) compared with the test and control line ratio (T/C ratio) of the BZ-nAb (r = −0.7089, p < 0.001). An almost perfect agreement was seen between the BZ-nAb and cPass sVNT results, with a strong negative correlation between the BZ-nAb T/C ratio and cPass sVNT percent inhibition (r = −0.8022, p < 0.001). In conclusion, the diagnostic performance of the BZ-nAb was comparable to that of the cPass sVNT, although the BZ-nAb had a slightly lower sensitivity.

scholarly journals Inference of SARS-CoV-2 spike-binding neutralizing antibody titers in sera from hospitalized COVID-19 patients by using commercial enzyme and chemiluminescent immunoassays

2020 ◽  
Author(s):  
Arantxa Valdivia ◽  
Ignacio Torres ◽  
Victor Latorre ◽  
Carla Frances-Gomez ◽  
Eliseo Albert ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Virus Neutralization ◽  
Pseudotyped Virus ◽  
S Protein ◽  
Neutralizing Activity ◽  
Virus Neutralization Assay ◽  
Degree Correlation ◽  
Antibody Titers

Background: Whether antibody levels measured by commercially-available enzyme or chemiluminescent immunoassays targeting the SARS-CoV-2 spike (S) protein can act as a proxy for serum neutralizing activity remains to be established for many of these assays. Objectives: To evaluate the degree of correlation between neutralizing antibodies (NtAb) binding the SARS-CoV-2 Spike (S) protein and SARS-CoV-2-S-IgG levels measured by four commercial immunoassays in sera drawn from hospitalized COVID-19 patients. Patients and Methods: Ninety sera from 51 hospitalized COVID-19 patients were assayed by a pseudotyped virus neutralization assay, the LIAISON SARS-CoV-2 S1/S2 IgG, the Euroimmun SARS-CoV-2 IgG ELISA, the MAGLUMI 2019-nCoV IgG and the COVID-19 ELISA IgG assays. Results: Overall, the results obtained with the COVID-19 ELISA IgG test showed the highest agreement with the NtAb assay (κ, 0.85; 95% CI, 0.63-1). The most sensitive tests were the pseudotyped virus NtAb assay and the COVID-19 ELISA IgG assay (92.2% for both). Overall, the degree correlation between antibody titers resulting in 50% virus neutralization (NtAb50) in the pseudotyped virus assay and SARS-CoV-2 IgG levels was strong for the Euroimmun SARS-CoV-2 IgG ELISA (Rho=0.73) and moderate for the remaining assays (Rho=0.48 to 0.59). The kinetic profile of serum NtAb50 titers could not be reliably predicted by any of the SARS-CoV-2 IgG immunoassays. Conclusions: the suitability of SARS-CoV-2-S-IgG commercial immunoassays for inferring neutralizing activity of sera from hospitalized COVID-19 patients varies widely across tests and is influenced by the time of sera collection after the onset of symptoms.

scholarly journals Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses

2020 ◽  
Vol 217 (11) ◽  
Author(s):  
Fabian Schmidt ◽  
Yiska Weisblum ◽  
Frauke Muecksch ◽  
Hans-Heinrich Hoffmann ◽  
Eleftherios Michailidis ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Activity ◽  
Human Monoclonal Antibodies ◽  
Neutralizing Activity ◽  
Immunodeficiency Virus ◽  
Vesicular Stomatitis ◽  
Hiv 1

The emergence of SARS-CoV-2 and the ensuing explosive epidemic of COVID-19 disease has generated a need for assays to rapidly and conveniently measure the antiviral activity of SARS-CoV-2–specific antibodies. Here, we describe a collection of approaches based on SARS-CoV-2 spike-pseudotyped, single-cycle, replication-defective human immunodeficiency virus type-1 (HIV-1), and vesicular stomatitis virus (VSV), as well as a replication-competent VSV/SARS-CoV-2 chimeric virus. While each surrogate virus exhibited subtle differences in the sensitivity with which neutralizing activity was detected, the neutralizing activity of both convalescent plasma and human monoclonal antibodies measured using each virus correlated quantitatively with neutralizing activity measured using an authentic SARS-CoV-2 neutralization assay. The assays described herein are adaptable to high throughput and are useful tools in the evaluation of serologic immunity conferred by vaccination or prior SARS-CoV-2 infection, as well as the potency of convalescent plasma or human monoclonal antibodies.

scholarly journals A Genetic System for Rhesus Monkey Rhadinovirus: Use of Recombinant Virus To Quantitate Antibody-Mediated Neutralization

2006 ◽  
Vol 80 (3) ◽  
pp. 1549-1562 ◽  
Author(s):  
John P. Bilello ◽  
Jennifer S. Morgan ◽  
Blossom Damania ◽  
Sabine M. Lang ◽  
Ronald C. Desrosiers
Keyword(s):  
B Cells ◽  
Rhesus Monkey ◽  
Rhesus Monkeys ◽  
Fluorescent Protein ◽  
Kaposi Sarcoma ◽  
Genetic System ◽  
Cell Types ◽  
Neutralization Assay ◽  
Neutralizing Activity ◽  
Virus Neutralization Assay

ABSTRACT Rhesus monkey rhadinovirus (RRV), a simian gamma-2 herpesvirus closely related to the Kaposi sarcoma-associated herpesvirus, replicates lytically in cultured rhesus monkey fibroblasts and establishes persistence in B cells. Overlapping cosmid clones were generated that encompass the entire 130-kilobase-pair genome of RRV strain 26-95, including the terminal repeat regions required for its replication. Cloned RRV that was produced by cotransfection of overlapping cosmids spanning the entire RRV26-95 genome replicated with growth kinetics and to titers similar to those of the parental, uncloned, wild-type RRV26-95. Expression cassettes for secreted-engineered alkaline phosphatase (SEAP) and green fluorescent protein (GFP) were inserted upstream of the R1 gene, and the cosmid-based system for RRV genome reconstitution was used to generate replication-competent, recombinant RRV that expressed either the SEAP or GFP reporter gene. Using the SEAP and GFP recombinant RRVs, assays were developed to monitor RRV infection, neutralization, and replication. Heat-inactivated sera from rhesus monkeys that were naturally or experimentally infected with RRV were assayed for their ability to neutralize RRV-SEAP and RRV-GFP infectivity using rhesus monkey fibroblasts. Sera from RRV-positive monkeys, but not RRV-negative monkeys, were consistently able to neutralize RRV infectivity when assayed by the production of SEAP activity or by the ability to express GFP. The neutralizing activity was present in the immunoglobulin fraction. Of the 17 rhesus monkeys tested, sera from rhesus monkey 26-95, i.e., the monkey that yielded the RRV 26-95 isolate, had the highest titer of neutralizing activity against RRV26-95. This cosmid-based genetic system and the reporter virus neutralization assay will facilitate study of the contribution of individual RRV glycoproteins to entry into different cell types, particularly fibroblasts and B cells.

scholarly journals A novel approach for evaluating antibodies against SARS-CoV-2 using menstrual blood collected from sanitary napkins before and after vaccination and evaluation of the visual napkin score

2021 ◽  
Author(s):  
Hiromitsu Shirasawa ◽  
Yukiyo Kumazawa ◽  
Shiori Kushima ◽  
Ayaka Fujishima ◽  
Wataru Sato ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Conflicts Of Interest ◽  
Antibody Test ◽  
Positive Control ◽  
University Hospital ◽  
Menstrual Blood ◽  
Blood Collection ◽  
Control Line ◽  
Sample Type ◽  
Antibody Testing

Abstract BackgroundAlthough saliva, whole blood, serum, plasma, urine, and feces have been used as specimens for SARS-CoV-2 antigen and antibody tests, menstrual blood has not been reported to date. Unlike invasive blood collection methods, menstrual blood collected non-invasively from participants can be used to evaluate the presence of antibodies against SARS-CoV-2. The purpose of our report is to show an association between menstrual blood and the presence of neutralizing antibodies acquired via mRNA vaccination and the usefulness of menstrual blood as a sample type for detecting SARS-CoV-2 antibodies, considering the volume of blood in sanitary napkins as visual napkin score (VNS).MethodsIn this study, we collected one napkin each from 40 participants visiting the outpatient gynecology clinic of our university hospital with no symptoms related to COVID-19 and attempted to collect their menstrual blood from the napkins. In 5 of 40 participants, menstrual blood was collected after at least one dose of mRNA vaccination. For this study, the maximum volume of menstrual blood collected was set as 980 μl. In addition, the classification of napkins based on the VNS was set, with level 1 being the lowest percentage of blood on the napkin (0–20%) and level 5 being the highest category (80–100%), according to the consensus of two researchers. We have evaluated used four different antibody testing kits using menstrual blood for detecting IgG and IgM.FindingsThe mean amount of menstrual blood collected from the 40 participants' sanitary napkins was 364 ± 372 μl; higher VNS indicated more menstrual blood collected. Statistically, VNS 3 or higher resulted in significantly higher menstrual blood collection than VNS 1 (p<0.01). With VNS 1, the collection of menstrual blood was complicated, and antibody test kits could not be tested for all eight participants. On the other hand, 31 of 32 participants (96.9%) with VNS 2 or higher could be tested with one or more antibody test kits. For all testing kits, 100% of tests with menstrual blood had a positive control line, and all participants who tested positive for IgG and IgM had received a COVID-19 mRNA vaccine. In the five participants after mRNA vaccination, only two of the four testing kits were all positive for IgG.InterpretationWe have, for the first time, evaluated antibodies against SARS-CoV-2 in menstrual blood collected from sanitary napkins with several antibody test kits. We found that if more than 20% of the napkin area has menstrual blood on it, sufficient menstrual blood can be collected for antibody testing. We also confirmed that menstrual blood collected from a sanitary napkin could be used to detect antibody after mRNA COVID-19 vaccination. We believe that our results are a pioneering effort that has not been reported previously and will lead to better public health and development of wearable devices.FundingThere are no conflicts of interest to disclose for this study.

scholarly journals Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses

2020 ◽  
Author(s):  
Fabian Schmidt ◽  
Yiska Weisblum ◽  
Frauke Muecksch ◽  
Hans-Heinrich Hoffmann ◽  
Eleftherios Michailidis ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Activity ◽  
Human Monoclonal Antibodies ◽  
Neutralizing Activity ◽  
Immunodeficiency Virus ◽  
Vesicular Stomatitis ◽  
Hiv 1

AbstractThe emergence of SARS-CoV-2 and the ensuing explosive epidemic of COVID19 disease has generated a need for assays to rapidly and conveniently measure the antiviral activity of SARS-CoV-2-specific antibodies. Here, we describe a collection of approaches based on SARS-CoV-2 spike-pseudotyped, single-cycle, replication-defective human immunodeficiency virus type-1 (HIV-1) and vesicular stomatitis virus (VSV), as well as a replication-competent VSV/SARS-CoV-2 chimeric virus. While each surrogate virus exhibited subtle differences in the sensitivity with which neutralizing activity was detected, the neutralizing activity of both convalescent plasma and human monoclonal antibodies measured using each virus correlated quantitatively with neutralizing activity measured using an authentic SARS-CoV-2 neutralization assay. The assays described herein are adaptable to high throughput and are useful tools in the evaluation of serologic immunity conferred by vaccination or prior SARS-CoV-2 infection, as well as the potency of convalescent plasma or human monoclonal antibodies.

scholarly journals mRNA-1273 and BNT162b2 mRNA vaccines have reduced neutralizing activity against the SARS-CoV-2 Omicron variant

2021 ◽  
Author(s):  
Venkata-Viswanadh Edara ◽  
Kelly E Manning ◽  
Madison Ellis ◽  
Lilin Lai ◽  
Kathryn M Moore ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Responses ◽  
Spike Protein ◽  
Live Virus ◽  
Neutralizing Activity ◽  
Virus Neutralization Assay ◽  
Fold Reduction

The BNT162b2 (Pfizer-BioNTech) and mRNA-1273 (Moderna) vaccines generate potent neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the global emergence of SARS-CoV-2 variants with mutations in the spike protein, the principal antigenic target of these vaccines, has raised concerns over the neutralizing activity of vaccine-induced antibody responses. The Omicron variant, which emerged in November 2021, consists of over 30 mutations within the spike protein. Here, we used an authentic live virus neutralization assay to examine the neutralizing activity of the SARS-CoV-2 Omicron variant against mRNA vaccine-induced antibody responses. Following the 2nd dose, we observed a 30-fold reduction in neutralizing activity against the omicron variant. Through six months after the 2nd dose, none of the sera from naive vaccinated subjects showed neutralizing activity against the Omicron variant. In contrast, recovered vaccinated individuals showed a 22-fold reduction with more than half of the subjects retaining neutralizing antibody responses. Following a booster shot (3rd dose), we observed a 14-fold reduction in neutralizing activity against the omicron variant and over 90% of boosted subjects showed neutralizing activity against the omicron variant. These findings show that a 3rd dose is required to provide robust neutralizing antibody responses against the Omicron variant.

scholarly journals A hemagglutination-based, semi-quantitative test for point-of-care determination of SARS-CoV-2 antibody levels

2021 ◽  
Author(s):  
Robert L. Kruse ◽  
Yuting Huang ◽  
Alyssa Lee ◽  
Xianming Zhu ◽  
Ruchee Shrestha ◽  
...  
Keyword(s):  
Point Of Care ◽  
Neutralizing Antibody ◽  
Antibody Test ◽  
Quantitative Information ◽  
Elisa Test ◽  
Lateral Flow ◽  
Plasma Samples ◽  
Hemagglutination Test ◽  
Lateral Flow Assays

Serologic, point-of-care tests to detect antibodies against SARS-CoV-2 are an important tool in the COVID-19 pandemic. The majority of current point-of-care antibody tests developed for SARS-CoV-2 rely on lateral flow assays, but these do not offer quantitative information. To address this, we developed a novel antibody test leveraging hemagglutination, employing a dry card format currently used for typing ABO blood groups. 200 COVID-19 patient and 200 control plasma samples were reconstituted with O-negative RBCs to form whole blood and added to dried viral-antibody fusion protein, followed by a stirring step and a tilting step, 3-minute incubation, and a second tilting step. The sensitivity for the hemagglutination test, Euroimmun IgG ELISA test and RBD-based CoronaChek lateral flow assay was 87.0%, 86.5%, and 84.5%, respectively, using samples obtained from recovered COVID-19 individuals. Testing pre-pandemic samples, the hemagglutination test had a specificity of 95.5%, compared to 97.3% and 98.9% for the ELISA and CoronaChek, respectively. A distribution of agglutination strengths was observed in COVID-19 convalescent plasma samples, with the highest agglutination score (4) exhibiting significantly higher neutralizing antibody titers than weak positives (2) (p<0.0001). Strong agglutinations were observed within 1 minute of testing, and this shorter assay time also increased specificity to 98.5%. In conclusion, we developed a novel rapid, point-of-care RBC agglutination test for the detection of SARS-CoV-2 antibodies that can yield semi-quantitative information on neutralizing antibody titer in patients. The five-minute test may find use in determination of serostatus prior to vaccination, post-vaccination surveillance and travel screening.

DETEKSI ANTIBODI MULTIPEL HEPATITIS C DALAM DARAH DONOR

2016 ◽  
Vol 21 (3) ◽  
pp. 261
Author(s):  
Ranti Permatasari ◽  
Aryati Aryati ◽  
Budi Arifah
Keyword(s):  
Hepatitis C ◽  
Predictive Value ◽  
Core Protein ◽  
Rapid Test ◽  
Antibody Test ◽  
Diagnostic Value ◽  
Hcv Infection ◽  
Antibody Detection ◽  
Hcv Rna ◽  
Hcv Antibody

Hepatitis C (HCV) infection could be spread by blood transfusion. Screening of HCV in donor blood could prevent HCV infection to the recipient. HCV antibody test using rapid test of multiple antibody detection by immunochromatography method is an easy and rapid test that could detect four HCV antibodies separately. The aim of this study was to evaluate the diagnostic value of antibody HCV using multiple antibody detection rapid test in diagnosing HCV infection. This was an analytical observational study with a cross sectional design. The samples consisted of 42 donors’ blood serum from the Surabaya Branch of the Indonesian Red Cross which underwent HCV infection test using ELISA method. The samples were then tested using PCR HCV RNA as the gold standard and antibody HCV multiple antibodydetection rapid test The diagnostic value of HCV antibody test using multiple antibody detection rapid test by immunochromatography method showed a diagnostic sensitivity of 100%, diagnostic specificity of 75%, positive predictive value of 66.7% and negative predictive value of 100%, a diagnostic efficiency of 83.3%, with a positive probability ratio of 4 times. The most often positive antibody pattern was four (4) positive antibodies (core protein, NS3, NS4 and NS5). Core protein (CP) and NS3 were the most often positive antibodies. Based on this study result, the HCV antibody test using multiple antibody detection rapid test by immunochromatography method has a good diagnostic value.

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