Cell-Culture Virus-Neutralization Test and Enzyme-Linked Immunosorbent Assay for Evaluation of Immunity in Chickens against Fowlpox

1985 ◽  
Vol 29 (3) ◽  
pp. 672 ◽  
Author(s):  
C. Buscaglia ◽  
R. A. Bankowski ◽  
L. Miers
Keyword(s):  
Cell Culture ◽  
Immunosorbent Assay ◽  
Neutralization Test ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Culture Virus

scholarly journals Evaluation of six commercial SARS-CoV-2 serology assays detecting different antibodies for clinical testing and serosurveillance

2021 ◽  
Author(s):  
Suellen Nicholson ◽  
Theo Karapanagiotidis ◽  
Arseniy Khvorov ◽  
Celia Douros ◽  
Francesca Mordant ◽  
...  
Keyword(s):  
Cell Culture ◽  
Neutralization Test ◽  
Serological Test ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Binding Assays ◽  
Humoral Immune ◽  
A Cell

Abstract Background Serological testing for SARS-CoV-2 complements nucleic acid tests for patient diagnosis and enables monitoring of population susceptibility to inform the COVID-19 pandemic response. It is important to understand the reliability of assays with different antigen or antibody targets to detect humoral immunity after SARS-CoV-2 infection and to understand how antibody (Ab) binding assays compare to those detecting neutralizing antibody (nAb), particularly as we move into the era of vaccines. Methods We evaluated the performance of six commercially available Enzyme-linked Immunosorbent Assays (ELISAs), including a surrogate virus neutralization test (sVNT), for detection of SARS-CoV-2 immunoglobulins (IgA, IgM, IgG), total or nAb. A result subset was compared to a cell culture-based microneutralisation (MN) assay. We tested sera from patients with prior RT-PCR confirmed SARS-CoV-2 infection, pre-pandemic sera and potential cross-reactive sera from patients with other non-COVID-19 acute infections. Results For sera collected > 14 days post-symptom onset, the assay achieving the highest sensitivity was the Wantai total Ab at 100% (95% confidence interval: 94.6-100) followed by 93.1% for Euroimmun NCP-IgG, 93.1% for GenScript sVNT, 90.3% for Euroimmun S1-IgG, 88.9% for Euroimmun S1-IgA and 83.3% for Wantai IgM. Specificity for the best performing assay was 99.5% for the Wantai total Ab and for the lowest performing assay was 97.1% for sVNT (as per IFU). The Wantai Total Ab had the best agreement with MN at 98% followed by Euroimmun S1-IgA, Euro NCP-IgG and sVNT (as per IFU) with (97%, 97% and 95% respectively) and Wantai IgM having the poorest agreement at 93%. Conclusion Performance characteristics of the SARS-CoV-2 serology assays detecting different antibody types are consistent with those found in previously published reports. Evaluation of the surrogate virus neutralization test in comparison to the Ab binding assays and a cell culture-based neutralization assay showed good result correlation between all assays. However correlation between the cell-based neutralization test and some assays detecting Ab’s not specifically involved in neutralization was higher than with the sVNT. This study demonstrates the reliability of different assays to detect the humoral immune response following SARS-CoV-2 infection, which can be used to optimise serological test algorithms for assessing antibody responses post SARS-CoV-2 infection or vaccination.

scholarly journals Evaluation of a Novel Reporter Virus Neutralization Test for Serological Diagnosis of Zika and Dengue Virus Infection

2017 ◽  
Vol 55 (10) ◽  
pp. 3028-3036 ◽  
Author(s):  
Chao Shan ◽  
Daniel A. Ortiz ◽  
Yujiao Yang ◽  
Susan J. Wong ◽  
Laura D. Kramer ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Reference Method ◽  
Laboratory Diagnosis ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reporter Virus ◽  
Screening Assay

ABSTRACT Currently, the laboratory diagnosis of Zika virus (ZIKV) infection is primarily through the detection of ZIKV RNA or antibodies against ZIKV proteins. The detection of viral RNA is highly sensitive and specific, but periods of viremia and viruria are brief, limiting the utility of ZIKV RNA assays. Instead, most ZIKV infections are diagnosed serologically, using an IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for screening, followed by a confirmatory plaque reduction neutralization test (PRNT). Typical turnaround times vary, due to assay incubation periods and a lack of clinical laboratories performing these tests. Recently, a novel luciferase-ZIKV- and -dengue virus (DENV)-based serological assay, which considerably improves the turnaround times and throughput for ZIKV diagnosis, was described. Using the traditional PRNT as a reference method, we evaluated the performance characteristics of the reporter virus neutralization test (RVNT) with 258 clinical serum specimens. The ZIKV RVNT produced primary ZIKV screening and secondary confirmation results in 4 days, with 100% reproducibility. As a screening assay, the ZIKV RVNT displayed excellent diagnostic accuracy, sensitivity, and specificity of 98.2%, 100%, and 98.1%, respectively. As a confirmatory assay, the ZIKV RVNT titers displayed 93.1% agreement with the traditional ZIKV PRNT titers. Overall, the RVNT accurately and reliably detects neutralizing antibodies in patient serum specimens, with improved turnaround times, and can be used for the serological detection of ZIKV infections. Due to the homogeneous 96-well format, the RVNT has also significantly improved the assay throughput to allow testing of a large number of specimens in a single run.

scholarly journals Comparison of a Monoclonal Antibody Capture ELISA (MACELISA) to Indirect ELISA and Virus Neutralization Test for the Serodiagnosis of Transmissible Gastroenteritis Virus

1993 ◽  
Vol 5 (1) ◽  
pp. 21-25 ◽  
Author(s):  
Ignacio Lanza ◽  
Pedro Rubio ◽  
Maria Mufioz ◽  
Pedro Ckmenes
Keyword(s):  
Indirect Elisa ◽  
Neutralization Test ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transmissible Gastroenteritis Virus ◽  
Gastroenteritis Virus ◽  
Infected Cells ◽  
Transmissible Gastroenteritis ◽  
Sensitivity Specificity

An enzyme-linked immunosorbent assay (ELISA) in which the antigen is captured to the plate by monoclonal antibodies (MACELISA) was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). The viral antigen was semipurified from TGEV-infected cells by simple ultracentrifugation. MACELISA results with 258 field sera were compared with those of a standard indirect ELISA and with the virus neutralization test (VNT). Sensitivity, specificity, and kappa values of MACELISA indicated a strong correlation with VNT results, whereas an indirect ELISA was less sensitive and much less specific than VNT. The serologic response of 4 pigs orally inoculated and intraperitoneally boostered with TGEV was compared using the 3 tests. Its sensitivity, specificity, and ability to use unpurified antigen make the MACELISA the advisable first step in TGEV serodiagnosis.

scholarly journals Evaluation of Serological Tests for Detection of Antibodies against Lumpy Skin Disease Virus

2020 ◽  
Vol 58 (9) ◽  
Author(s):  
Nina Krešić ◽  
Ivana Šimić ◽  
Tomislav Bedeković ◽  
Žaklin Acinger-Rogić ◽  
Ivana Lojkić
Keyword(s):  
Skin Disease ◽  
Neutralizing Antibodies ◽  
Large Scale ◽  
Neutralization Test ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Surveillance Program ◽  
Kappa Index ◽  
Lumpy Skin Disease

ABSTRACT Lumpy skin disease (LSD) is an emerging, transboundary viral pox disease affecting cattle of all ages and breeds. The serological assay for monitoring immunity following vaccination is a virus neutralization test (VNT/OIE) that determines the neutralization index (NI). The first validated enzyme-linked immunosorbent assay (ELISA; IDVet) has become commercially available, facilitating large-scale serosurveillance for LSD. Although the VNT is labor intensive and time consuming, it is still the recommended test by the OIE. Thus, in this study, we modified the virus neutralization test by employing Madin-Darby bovine kidney (MDBK) cells. The qualitative results obtained with the modified method were compared to the qualitative results obtained by VNT/OIE and ELISA. We used blood sera received within a surveillance program for LSD in 2018. In total, 291 serum samples were tested using VNT/MDBK and ELISA. Of 291 samples, 80 samples were tested by VNT/OIE and used for comparison of the performances between VNT/MDBK and VNT/OIE. The compatibility of results obtained by VNT/MDBK and VNT/OIE resulted in a kappa index of 0.9 with overall proportion agreement of 0.96. Agreement between VNT/MDBK and VNT/OIE was achieved in 56 positive and 21 negative samples. The compatibility of results obtained by ELISA and VNT/MDBK were compared on 291 samples in total and resulted in a kappa index 0.834 with overall proportion agreement of 0.955. Agreement between ELISA and VNT/MDBK was achieved in 238 positive and 40 negative samples. The results obtained demonstrated a strong correlation between VNT/MDBK and the other two methods, indicating the suitability of VNT/MDBK for the detection of the LSD virus-specific neutralizing antibodies.

scholarly journals Serological Survey for Foot-and-Mouth Disease Virus in Wildlife in Eastern Africa and Estimation of Test Parameters of a Nonstructural Protein Enzyme-Linked Immunosorbent Assay for Buffalo

2008 ◽  
Vol 15 (6) ◽  
pp. 1003-1011 ◽  
Author(s):  
B. M. D. C. Bronsvoort ◽  
S. Parida ◽  
I. Handel ◽  
S. McFarland ◽  
L. Fleming ◽  
...  
Keyword(s):  
Disease Virus ◽  
Immunosorbent Assay ◽  
Neutralization Test ◽  
Nonstructural Protein ◽  
Foot And Mouth Disease ◽  
Virus Neutralization Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Parameters ◽  
Mouth Disease Virus ◽  
Protein Enzyme

ABSTRACT In this study we estimate the seroprevalence of foot-and-mouth disease virus (FMDV) in wildlife from eastern and central Africa. Sera were sourced from between 1994 and 2002 from a rinderpest surveillance program. Our study compared a nonstructural protein enzyme-linked immunosorbent assay (Cedi test) with a virus neutralization test. The study shows that there is only a low seroprevalence of FMDV in sampled nonbuffalo species. The seroprevalence in the Cape buffalo was high for SAT2, lower for SAT1, and lowest for SAT3. As the SAT2 serotype was most prevalent, the Cedi test largely reflected the occurrence of SAT2-positive animals. The results also suggest that SAT2 became dominant around 1998, with a large increase in seroprevalence. The sensitivity and specificity of the Cedi test were estimated by comparison to the combined virus neutralization test results from all three SAT tests. A Bayesian implementation of the Hui-Walter latent class model was used to estimate the test parameters. The model permits estimation in the absence of a gold standard test. The final model, using noninformative priors and assuming conditional independence of test performance, estimated Cedi test sensitivity at 87.7% and specificity at 87.3%. These estimates are similar to those for domestic bovines; they suggest that the Cedi test is a useful tool for screening buffalo for infection with the various serotypes of FMDV.

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