scholarly journals A high-throughput neutralization assay for yellow fever serodiagnostics

2021 ◽  
Author(s):  
Madina Rasulova ◽  
Thomas Vercruysse ◽  
Jasmine Paulissen ◽  
Catherina Coun ◽  
Vanessa Suin ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Yellow Fever ◽  
High Throughput ◽  
Gold Standard ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
High Sensitivity ◽  
Neutralization Assay ◽  
Differential Diagnostics ◽  
Clinical Use

Quick and accurate detection of neutralizing antibodies (nAbs) against yellow fever is essential in serodiagnosis during outbreaks, for surveillance and to evaluate vaccine efficacy in population-wide studies. All this requires serological assays that can process a large number of samples in a highly standardized format. Albeit being laborious, time-consuming and limited in throughput, classical plaque reduction neutralization test (PRNT) is still considered gold standard for the detection and quantification of nAbs due to its sensitivity and specificity. Here we report the development of an alternative fluorescence-based serological assay (SNTFLUO) with an equally high sensitivity and specificity that is fit for high-throughput testing with the potential for automation. Finally, our novel SNTFLUO was cross-validated in several reference laboratories and against international WHO standards showing its potential to be implemented in clinical use. SNTFLUO assays with similar performance are available for the Japanese encephalitis, Zika and dengue viruses amenable for differential diagnostics.

scholarly journals Performance of three SARS-CoV-2 immunoassays, three rapid lateral flow tests and a novel bead-based affinity surrogate test for the detection of SARS-CoV-2 antibodies in human serum

2021 ◽  
Author(s):  
Manuel Krone ◽  
Julia Gütling ◽  
Johannes Wagener ◽  
Thiên-Trí Lâm ◽  
Christoph Schoen ◽  
...  
Keyword(s):  
Human Serum ◽  
Sensitivity And Specificity ◽  
Symptom Onset ◽  
Gold Standard ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Lateral Flow ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Lateral Flow Tests

For the control of immunity in COVID-19 survivors and vaccinated subjects there is an urgent need for reliable and rapid serological assays. Based on samples from 63 COVID-19 survivors up to seven months after symptom onset, and on 50 serum samples taken before the beginning of the pandemic, we compared the performance of three commercial immunoassays for the detection of SARS-CoV-2 IgA and IgG antibodies (Euroimmun SARS-COV-2 IgA/IgG, Mikrogen recomWell SARS-CoV-2 IgA/IgG, and SERION ELISA agile SARS-CoV-2 IgA/IgG) and three rapid lateral flow (immunochromatographic) tests (Abbott Panbio COVID-19 IgG/IgM, NADAL COVID-19 IgG/IgM, and Cleartest Corona 2019-nCOV IgG/IgM) with a plaque-reduction neutralization test (PRNT50) representing the gold standard. Fifty-seven out of 63 PCR-confirmed COVID-19 patients (90%) showed neutralizing antibodies. The sensitivity of the seven assays ranged from 7.0% to 98.3%, the specificity from 86.0% to 100.0%. Only one commercial immunoassay showed a sensitivity and specificity of greater than 98%.

scholarly journals A high-throughput neutralizing antibody assay for COVID-19 diagnosis and vaccine evaluation

2020 ◽  
Author(s):  
Antonio E. Muruato ◽  
Camila R. Fontes-Garfias ◽  
Ping Ren ◽  
Mariano A. Garcia-Blanco ◽  
Vineet D. Menachery ◽  
...  
Keyword(s):  
High Throughput ◽  
Gold Standard ◽  
Neutralizing Antibodies ◽  
Vaccine Development ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Assay ◽  
Plasma Therapy ◽  
High Throughput Assay ◽  
Key Parameter

AbstractVirus neutralization remains the gold standard for determining antibody efficacy. Therefore, a high-throughput assay to measure SARS-CoV-2 neutralizing antibodies is urgently needed for COVID-19 serodiagnosis, convalescent plasma therapy, and vaccine development. Here we report on a fluorescence-based SARS-CoV-2 neutralization assay that detects SARS-CoV-2 neutralizing antibodies in COVID-19 patient specimens and yields comparable results to plaque reduction neutralizing assay, the gold standard of serological testing. Our approach offers a rapid platform that can be scaled to screen people for antibody protection from COVID-19, a key parameter necessary to safely reopen local communities.

Development of a high-throughput β-Gal-based neutralization assay for quantitation of herpes simplex virus-neutralizing antibodies in human samples

Vaccine ◽  
2016 ◽  
Vol 34 (33) ◽  
pp. 3901-3906 ◽  
Author(s):  
Amy Baccari ◽  
Michael Cooney ◽  
Tamara P. Blevins ◽  
Lynda A. Morrison ◽  
Shane Larson ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
High Throughput ◽  
Neutralizing Antibodies ◽  
Neutralization Assay ◽  
Human Samples ◽  
Simplex Virus

scholarly journals Neutralization Assay Using a Modified Vaccinia Virus Ankara Vector Expressing the Green Fluorescent Protein Is a High-Throughput Method To Monitor the Humoral Immune Response against Vaccinia Virus

2004 ◽  
Vol 11 (2) ◽  
pp. 406-410 ◽  
Author(s):  
Antonio Cosma ◽  
Silja Bühler ◽  
Rashmi Nagaraj ◽  
Caroline Staib ◽  
Anna-Lena Hammarin ◽  
...  
Keyword(s):  
Immune Response ◽  
Green Fluorescent Protein ◽  
Vaccinia Virus ◽  
High Throughput ◽  
Neutralizing Antibodies ◽  
Fluorescent Protein ◽  
Safety Concern ◽  
Neutralization Assay ◽  
Modified Vaccinia Virus Ankara ◽  
Green Fluorescent

ABSTRACT Vaccination against smallpox is again considered in order to face a possible bioterrorist threat, but the nature and the level of the immune response needed to protect a person from smallpox after vaccination are not totally understood. Therefore, simple, rapid, and accurate assays to evaluate the immune response to vaccinia virus need to be developed. Neutralization assays are usually considered good predictors of vaccine efficacy and more informative with regard to protection than binding assays. Currently, the presence of neutralizing antibodies to vaccinia virus is measured using a plaque reduction neutralization test, but this method is time-consuming and labor-intensive and has a subjective readout. Here, we describe an innovative neutralization assay based on a modified vaccinia virus Ankara (MVA) vector expressing the green fluorescent protein (MVA-gfp). This MVA-gfp neutralization assay is rapid and sensitive and has a high-throughput potential. Thus, it is suitable to monitor the immune response and eventually the efficacy of a large campaign of vaccination against smallpox and to study the vector-specific immune response in clinical trials that use genetically engineered vaccinia viruses. Most importantly, application of the highly attenuated MVA eliminates the safety concern in using the replication-competent vaccinia virus in the standard clinical laboratory.

scholarly journals Toxin Detection by Cell Culture Neutralization Assay [CYT] and Toxin based EIA [Tox EIA] among Recurrent Episodes of CDI Diagnosed by PCR

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S395-S395
Author(s):  
Mini Kamboj ◽  
Tracy Mcmillen ◽  
Hoi Yan Chow ◽  
Jennifer Brite ◽  
N Esther Babady
Keyword(s):  
Gold Standard ◽  
Diagnostic Tests ◽  
Ad Hoc ◽  
Index Case ◽  
Extended Period ◽  
High Sensitivity ◽  
Neutralization Assay ◽  
Significant Implication ◽  
Stool Samples ◽  
Positive Recurrent

Abstract Background The Ad Hoc C. difficile surveillance working group defines recurrent C. difficile infection as a second episode occurring >8 weeks after the index case. Due to its high sensitivity, diagnosis of recurrent CDI by PCR is extremely challenging in patients who may have persistent, PCR detectable shedding of toxigenic C. difficile (TCD) for an extended period of time after treatment of the initial CDI episode. CYT, which detects C. difficile toxin antigen, is a cumbersome test to perform but is considered as the current clinical diagnostic gold standard for CDI diagnosis. Aim: To determine the CYT and Toxin A/B EIA positivity among patients with recurrent CDI episodes detected by PCR. We further characterized the performance of diagnostic tests based on whether the recurrent episode was a relapse or reinfection. Methods During a three month study period, CYT and Tox A/B EIA was performed on consecutive stool samples submitted from PCR positive recurrent episodes of CDI. For the purpose of this study, recurrence was defined as a second episode of CDI that occurred within 120 days from the most recent episode. MLST analysis was performed as previously described to characterize relapse and reinfection among the recurrent episodes (2). Results Thirty-five recurrent episodes occurred over the study period. 21/35 [60%] were positive by CYT and 12/35 [34%] by Tox A/B EIA. Among the recurrent CDI episodes, 16 (46%) were genotypical confirmed as relapse with the original infecting strain. Majority of these relapses were positive by CYT (81%) when compared with Tox EIA (43%). Among patients with geno typically confirmed reinfection (n = 8), CYT and EIA positivity was 63 % and 50 % respectively. For the remaining 11 episodes, TCD was not retrievable in culture, CYT and EIA positivity among this group was 27 % and 9 % respectively. Conclusion Forty percent of recurrent CDI episodes detected by PCR could not be confirmed by CYT. EIA missed 66 % of CYT positive recurrent CDI. The performance of CYT and EIA varied among recurrences due to relapse and reinfection. These results have significant implication for reporting of CDI HAI rates. Disclosures All authors: No reported disclosures.

scholarly journals Real-time RT-PCR diagnostics of virus causing COVID-19

2020 ◽  
Vol 13 (1) ◽  
pp. 52-63 ◽  
Author(s):  
E. V. Goncharova ◽  
A. E. Donnikov ◽  
V. V. Kadochnikova ◽  
S. A. Morozova ◽  
M. N. Boldyreva ◽  
...  
Keyword(s):  
Real Time ◽  
Sensitivity And Specificity ◽  
High Sensitivity ◽  
Medical Product ◽  
World Health ◽  
Differential Diagnostics ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Study Results ◽  
Pcr Diagnostics

Aim: the study was aimed to develop a reagent kit for the real-time RT-PCR diagnostics of virus causing COVID-19.Materials and Methods. Three target sites were chosen in the genome SARS-CoV-2. The testing included 220 samples, 48 artificially created positive samples (made from patients’ biomaterial) and 172 clinical samples (scrapes from nasal and pharyngeal cavities, bronchoalveolar lavage, expectoration, endotracheal/nasopharyngeal aspirate, feces, post-mortem material), obtained from two medical centers. Preliminary, the obtained biomaterial was analyzed with a reagent kit of comparison. The evaluation was performed with a confidential interval CI 95%. The calculation of CI for the sensitivity and specificity was made based on the distribution of χ2.Results. The authors developed a technology of novel coronavirus infection (COVID-19) real-time RT-PCR diagnostics for the application in practical healthcare and proposed the variants of testing at all the stages (preanalytical, analytical, and post-analytical, including automated results processing). The proposed reagent kit meets the requirements of the World Health Organization and the Ministry of Healthcare of the Russian Federation. The study results demonstrated high sensitivity and specificity. The sensitivity was 100% (95% CI) 95.6–100%; the specificity was 100% (95% CI) 96.7–100%.Conclusion. The proposed reagent kit was registered in the RF as a medical product; the registration certificate No. RZN 2020/9948 dated 01.04.2020. The application of the reagent kit in network laboratories will provide patients with access to testing for the virus causing COVID-19 and contribute to quick differential diagnostics, improvement of pandemic control, and accurate statistics on the spread of the virus. 

scholarly journals Protective levels of canine distemper virus antibody in an urban dog population using plaque reduction neutralization test

2004 ◽  
Vol 71 (3) ◽  
Author(s):  
O.I. Oyedele ◽  
D.O. Oluwayelu ◽  
S.I.B. Cadmus ◽  
F.D. Adu
Keyword(s):  
Neutralizing Antibodies ◽  
Canine Distemper Virus ◽  
Neutralization Test ◽  
Neutralization Assay ◽  
Canine Distemper ◽  
Virus Antibody ◽  
Post Vaccination ◽  
Blood Samples ◽  
Highly Sensitive ◽  
Positive Serum

Blood samples from 50 dogs were collected at three veterinary clinics in Ibadan and Abuja, Nigeria and the serum from each sample was evaluated serologically for neutralizing antibodies against canine distemper virus (CDV) by the highly sensitive plaque reduction (PRN) neutralization assay. Thirteen dogs had plaque reduction neutralization titres of 0-100, seven had titres of 100-1 000 while 30 had titres ranging from 1 000-6 000. The PRN titres of vaccinated dogs were found to be significantly higher than unvaccinated dogs. The widespread use of the highly reproducible PRN test for the evaluation of antibody response to CDV may be very important in the generation of international CDV positive serum standards that should help to improve pre-and post-vaccination testing of dogs worldwide.

Diagnostic efficacy of joint ultrasonography, dual-energy computed tomography and minimally invasive arthroscopy on knee gouty arthritis, a comparative study

2021 ◽  
pp. 20200493
Author(s):  
Yuesheng Xie ◽  
Ling Li ◽  
Riqiang Luo ◽  
Ting Xu ◽  
Lin Yang ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Knee Joint ◽  
Minimally Invasive ◽  
Sensitivity And Specificity ◽  
Gold Standard ◽  
Diagnostic Performance ◽  
Gouty Arthritis ◽  
High Sensitivity ◽  
Dual Energy ◽  
Double Contour

Objective: This study aimed to investigate the diagnostic performance of minimally invasive arthroscopy for knee gout when comparing with joint ultrasonography and dual-energy computed tomography (DECT). Methods: From January 2016 to December 2018, 121 inpatients with knee joint swelling and pain were prospectively enrolled, including 63 gout patients and 58 non-gout patients. All patients underwent pre-operative ultrasonography and DECT to evaluate knee joint monosodium urate (MSU) deposits, followed by minimally invasive arthroscopy. The gold-standard for gout diagnosis was defined as the detection of MSU crystals in the synovial fluid under polarizing microscopic or pathological analysis. Results: The diagnostic results of ultrasonic double contour sign, hyperechogenic foci, MSU deposition (detected by DECT), MSU deposition (detected by arthroscopy) and MSU deposition in cartilage (detected by arthroscopy) were significantly associated with that of the gold-standard. Except for hyperechogenic foci, the other four indexes had high sensitivity and specificity (approximately or over 80%) and a large odds ratio (OR) (14.73 to 36.56), indicating good diagnostic performance. Detection of MSU deposition in cartilage by arthroscopy had a good diagnostic agreement with the ultrasonic double contour sign (κ = 0.711, p < 0.001). Conclusion: Joint ultrasonography, DECT, and minimally invasive arthroscopy had high sensitivity and specificity for the diagnosis of knee gouty arthritis. Minimally invasive arthroscopy was superior to joint ultrasonography and DECT, which can be a useful supplement for the diagnosis of gout. Advances in knowledge: This is the first study comparing the diagnostic performance for knee gout among the joint ultrasonography, DECT, and minimally invasive arthroscopy.

scholarly journals Development of a Sensitive Detection Method for Alphaviruses and Its Use as a Virus Neutralization Assay

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1191
Author(s):  
Christin Schmidt ◽  
Mario Perkovic ◽  
Barbara S. Schnierle
Keyword(s):  
Neutralizing Antibodies ◽  
High Sensitivity ◽  
Neutralization Assay ◽  
Open Reading Frames ◽  
Virus Neutralization ◽  
Structural Genes ◽  
Virus Neutralization Assay ◽  
Subgenomic Promoter ◽  
Mayaro Virus ◽  
Reading Frames

Alphaviruses have a single-stranded, positive-sense RNA genome that contains two open reading frames encoding either the non-structural or the structural genes. Upon infection, the genomic RNA is translated into the non-structural proteins (nsPs). NsPs are required for viral RNA replication and transcription driven from the subgenomic promoter (sgP). Transfection of an RNA encoding the luciferase gene under the control of the sgP into cells enabled the detection of replication-competent chikungunya virus (CHIKV) or Mayaro virus (MAYV) with high sensitivity as a function of the induced luciferase activity. This assay principle was additionally used to analyze virus-neutralizing antibodies in sera and might be an alternative to standard virus neutralization assays based on virus titration or the use of genetically modified tagged viruses.

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