scholarly journals Sensitivity of the biotin-avidin-peroxidase (BAP) technique for rabies virus neutralizing antibody assay and the measurement of neutralizing antibodies of vaccinated human sera.

1983 ◽  
Vol 11 (3/4) ◽  
pp. 217-224
Author(s):  
AKEHISA SHICHIJO ◽  
KUMATO MIFUNE ◽  
KUNIAKI SAKAMOTO ◽  
AKIRA YAMADA
Keyword(s):  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Antibody Assay ◽  
Human Sera

scholarly journals Probing neutralizing-antibody responses against emerging measles viruses (MVs): immune selection of MV by H protein-specific antibodies?

2005 ◽  
Vol 86 (2) ◽  
pp. 365-374 ◽  
Author(s):  
Sabine Santibanez ◽  
Stefan Niewiesk ◽  
Alla Heider ◽  
Jürgen Schneider-Schaulies ◽  
Guy A. M. Berbers ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Protein A ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Neutralizing Capacity ◽  
Human Sera ◽  
H Protein ◽  
Neutralizing Epitopes ◽  
Selection Of

Measles virus (MV) infection and vaccination induce long-lasting immunity and neutralizing-antibody responses that are directed against the MV haemagglutinin (H) and the fusion (F) protein. A new MV genotype, D7, emerged recently in western Germany and rapidly replaced the long-term endemically circulating genotypes C2 and D6. Analysis of the H gene of C2, D6, D7 and vaccine viruses revealed uniform sequences for each genotype. Interestingly, a consistent exchange of seven distinct amino acids in the D7 H was observed when compared with residues shared between C2, D6 and vaccine viruses, and one exchange (D416→N) in the D7 H was associated with an additional N-linked glycosylation. In contrast, the F gene is highly conserved between MVs of these genotypes. To test whether the D7 H protein escapes from antibody responses that were raised against earlier circulating or vaccine viruses, the neutralizing capacity of mAbs recognizing seven distinct domains on the H of an Edmonston-related MV was compared. The mAbs revealed a selective and complete loss of two neutralizing epitopes on the D7 H when compared with C2, D6 and vaccine viruses. To assess whether these alterations of the D7 H affect the neutralizing capacity of polyclonal B-cell responses, genotype-specific antisera were produced in cotton rats. However, no significant genotype-dependent difference was found. Likewise, human sera obtained from vaccinees (n=7) and convalescents (n=6) did not distinguish between the MV genotypes. Although the hypothesis of selection of D7 viruses by pre-existing neutralizing antibodies is compatible with the differing pattern of neutralizing epitopes on the H protein, it was not confirmed by the results of MV neutralization with polyclonal sera.

scholarly journals Evaluation of Rapid Neutralizing Antibody Detection Test against Rabies Virus in Human Sera

2015 ◽  
Vol 43 (2) ◽  
pp. 111-116 ◽  
Author(s):  
Kieu Anh Thi Nguyen ◽  
Thu Tuyet Nguyen ◽  
Dong Vinh Nguyen ◽  
Giang Chau Ngo ◽  
Cam Nhat Nguyen ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Neutralizing Antibody ◽  
Antibody Detection ◽  
Human Sera

scholarly journals Calculating rabies virus neutralizing antibodies titres by flow cytometry

2002 ◽  
Vol 44 (3) ◽  
pp. 151-154 ◽  
Author(s):  
Juliano BORDIGNON ◽  
Fabiano COMIN ◽  
Sílvia Córdoba P. FERREIRA ◽  
Graciane M. M. CAPORALE ◽  
José Hermênio Cavalcante LIMA FILHO ◽  
...  
Keyword(s):  
Cell Culture ◽  
Flow Cytometry ◽  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Standard Test ◽  
Serum Samples ◽  
Reference Serum ◽  
Infection Inhibition

The determination of the rabies neutralizing antibody (VNA) response after immunization against rabies is an acceptable index of the efficacy of a vaccine and a successful treatment. Several tests have been developed in attempt to improve the assessment of VNA, from mice inoculation to cell-culture fluorescence inhibition tests. All of them, however, present special difficulties in terms of reading or accuracy. The present study describes a neutralization test performed in cell-culture appraised by flow cytometry (FC). Serial dilutions of the serum samples were mixed in vitro with rabies virus before the addition of BHK-21 cells. After 24h-incubation, cells were released by trypsin treatment, fixed and permeabilized with a p-formaldehyde solution and stained with a rabies virus nucleocapsid protein-specific antibody conjugate. The percentage of virus infection inhibition caused by specific antibodies present in the serum were evaluated in a Beckton & Dickinson FACSCalibur® flow cytometer. A correlation curve between the IU/ml content and the percentage of infective inhibition was built with a reference serum and the VNA titers of serum samples were obtained by extrapolation. Titers obtained by FC and standard test showed an effective pairing results (p < 0.01), with a correlation coefficient (r) = 0.7. These results permit to envisage the FC as a suitable technique to evaluate VNA in sera from immunized animals and likely in human serum samples. Nevertheless, new studies comparing FC to gold-standard techniques are required for determining the FC values of Sensibility and Specificity .

Antibody production in goat milk serum after virus instillation of goat mammary gland. VII. Biochemical studies on sham infection with rabies and other non-propagating agents

1975 ◽  
Vol 21 (5) ◽  
pp. 655-661 ◽  
Author(s):  
Arthur E. Pasieka ◽  
L. F. Guerin ◽  
Chas. A. Mitchell
Keyword(s):  
Mammary Gland ◽  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Analytical Ultracentrifugation ◽  
Sedimentation Coefficient ◽  
Goat Milk ◽  
Neutralizing Antibody ◽  
Antibody Activity ◽  
Gel Chromatography ◽  
Milk Serum

This report describes the development of a method for the production and isolation of neutralizing antibodies against rabies virus (ERA strain) and other agents in the goat mammary gland during active lactation. The rabies virus did not propagate in the gland, but neutralizing antibodies were produced by serial instillations of the antigen. This process was termed 'sham infection.' Antibodies first appeared in the milk about the 20th day and the titer increased until the 28th day. The antibodies were found to be associated with the lactogammaglobulin. The milk globulins were fractionated by gel chromatography and the fraction containing the antibody was isolated. This fraction was further purified and characterized by immunodiffusion, immunoelectrophoresis, and an analytical ultracentrifugation technique. The neutralizing antibody activity after the 29th day was associated with the milk serum IgG fraction. This active fraction was found to have a sedimentation coefficient of 6.8 Svedberg units.

scholarly journals Chemiluminescence Immunoassay Based Serological Immunoassays for Detection of SARS-CoV-2 Neutralizing Antibodies in COVID-19 Convalescent Patients and Vaccinated Population

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1508
Author(s):  
Qiangling Yin ◽  
Yecheng Zhang ◽  
Lijun Lian ◽  
Yuanyuan Qu ◽  
Wei Wu ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Antibody Concentration ◽  
Inhibition Assay ◽  
Healthy Donors ◽  
Kappa Value ◽  
Human Sera ◽  
Highly Correlated

The development of rapid serological detection methods re urgently needed for determination of neutralizing antibodies in sera. In this study, four rapid methods (ACE2-RBD inhibition assay, S1-IgG detection, RBD-IgG detection, and N-IgG detection) were established and evaluated based on chemiluminescence technology. For the first time, a broadly neutralizing antibody with high affinity was used as a standard for the quantitative detection of SARS-CoV-2 specific neutralizing antibodies in human sera. Sera from COVID-19 convalescent patients (N = 119), vaccinated donors (N = 86), and healthy donors (N = 299) confirmed by microneutralization test (MNT) were used to evaluate the above methods. The result showed that the ACE2-RBD inhibition assay calculated with either ACE2-RBD binding inhibition percentage rate or ACE2-RBD inhibiting antibody concentration were strongly correlated with MNT (r ≥ 0.78, p < 0.0001) and also highly consistent with MNT (Kappa Value ≥ 0.94, p < 0.01). There was also a strong correlation between the two evaluation indices (r ≥ 0.99, p < 0.0001). Meanwhile, S1-IgG and RBD-IgG quantitative detection were also significantly correlated with MNT (r ≥ 0.73, p < 0.0001), and both methods were highly correlated with each other (r ≥ 0.95, p < 0.0001). However, the concentration of N-IgG antibodies showed a lower correlation with the MNT results (r < 0.49, p < 0.0001). The diagnostic assays presented here could be used for the evaluation of SARS-CoV-2 vaccine immunization effect and serological diagnosis of COVID-19 patients, and could also have guiding significance for establishing other rapid serological methods to surrogate neutralization tests for SARS-CoV-2.

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.

scholarly journals Antibodies against rabies virus in dogs with and without history of vaccination in Santa Maria - RS - Brazil

2017 ◽  
Vol 47 (11) ◽  
Author(s):  
Karina Gonzalez Fernandes ◽  
Mathias Martins ◽  
Bruna Portolan Amaral ◽  
Juliana Felipetto Cargnelutti ◽  
Rudi Weiblen ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
World Health ◽  
Serum Samples ◽  
Antibody Titers ◽  
Group A ◽  
Group B ◽  
Santa Maria ◽  
Rabies Vaccination

ABSTRACT: The present study investigated the frequency and magnitude of neutralizing antibodies to rabies virus (RABV) in dogs with and without historic of vaccination in Santa Maria/RS. Group A included serum samples from 440 dogs with recent historic of vaccination against rabies, obtained during the 2015 rabies vaccination campaign. Group B included 300 serum samples from dogs submitted to the Veterinary Hospital of the Universidade Federal de Santa Maria in 2015, whose historic of rabies vaccination was unknown. Serum samples were submitted to the rapid fluorescent focus inhibition test (RFFIT) to detect neutralizing antibodies against RABV. In group A, 70.6% (310/440) of the samples had neutralizing antibody titers ≥0.5 international units per milliliter (IU mL-1), considered an indicative of protection against rabies by the World Health Organization. However, approximately 30% of the dogs did not contain antibodies in adequate levels. In group B, 42.3% (127/300) of the samples contained neutralizing antibody titers ≥0.5IU mL-1 and 57.7% (173/300) were negative or contained titers below of the value considered immunized. These results demonstrate that an important proportion of vaccinated dogs (~30%) did not develop adequate antibody levels, mainly those receiving a single vaccine dose. Serologic testing of animals with unknown historic of vaccination revealed relatively low vaccine coverage in the general dog population. Thus, reformulation of immunization strategies - especially the recommendation of a boost vaccination 30 days after the primary dose - and extension of vaccination campaigns are necessary to reach adequate levels and coverage of immunity against RABV in the canine population.

scholarly journals Measurement of the Humoral Immune Response following an Incident Human Papillomavirus Type 16 or 18 Infection in Young Women by a Pseudovirion-Based Neutralizing Antibody Assay

2008 ◽  
Vol 15 (9) ◽  
pp. 1387-1390 ◽  
Author(s):  
Jane Steele ◽  
Stuart Collins ◽  
Kaisheng Wen ◽  
Gordon Ryan ◽  
Christothea Constandinou-Williams ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Young Women ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Antibody Assay ◽  
Hpv 16 ◽  
Hpv 18

ABSTRACT We have evaluated a neutralizing antibody assay which uses human papillomavirus (HPV) type 16 (HPV-16) and HPV-18 pseudovirions carrying a secretory alkaline phosphatase reporter gene and which can potentially measure functionally relevant HPV type-specific neutralizing antibodies. The reproducibility of the assay was excellent; for HPV-16, the intra- and interassay kappa values were 0.95 and 0.90, respectively; and for HPV-18, the corresponding values were 0.90 and 0.90. This assay was used to describe the kinetics of the neutralizing antibody response in a cohort of 42 young women who were recruited soon after first intercourse and who first tested positive for HPV-16 DNA or HPV-18 DNA, or both, during follow-up. Most women seroconverted following the first detection of type-specific HPV DNA and remained seropositive until the end of follow-up. Our findings are broadly consistent with those of two other cohort studies which have measured the serological response following an incident infection by using the technically simpler virus-like-particle-based enzyme-linked immunosorbent assay.

scholarly journals Immunogenicity Studies in Carnivores Using a Rabies Virus Construct with a Site-Directed Deletion in the Phosphoprotein

2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
Ad Vos ◽  
Karl-Klaus Conzelmann ◽  
Stefan Finke ◽  
Thomas Müller ◽  
Jens Teifke ◽  
...  
Keyword(s):  
Oral Dose ◽  
Rabies Virus ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Oral Vaccination ◽  
P Gene ◽  
Interferon Induction ◽  
Antibody Titres ◽  
A Site

Different approaches have been applied to develop highly attenuated rabies virus vaccines for oral vaccination of mesocarnivores. One prototype vaccine construct is SAD dIND1, which contains a deletion in the P-gene severely limiting the inhibition of type-1 interferon induction. Immunogenicity studies in foxes and skunks were undertaken to investigate whether this highly attenuated vaccine would be more immunogenic than the parental SAD B19 vaccine strain. In foxes, it was demonstrated that SAD dIND1 protected the animals against a rabies infection after a single oral dose, although virus neutralizing antibody titres were lower than in foxes orally vaccinated with the SAD B19 virus as observed in previous experiments. In contrast, skunks receiving 107.5FFU SAD dIND1 did not develop virus neutralizing antibodies and were not protected against a subsequent rabies infection.

scholarly journals Development and Optimization of a Novel Assay To Measure Neutralizing Antibodies against Clostridium difficile Toxins

2013 ◽  
Vol 20 (4) ◽  
pp. 517-525 ◽  
Author(s):  
Jinfu Xie ◽  
Julie Zorman ◽  
Lani Indrawati ◽  
Melanie Horton ◽  
Keri Soring ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Vero Cells ◽  
Seeding Density ◽  
Content Type ◽  
Human Sera ◽  
Antibody Titers ◽  
Microscopic Inspection ◽  
Antibody Levels

ABSTRACTClostridium difficileproduces two major virulence toxins, toxin A (TcdA) and toxin B (TcdB). Antitoxin antibodies, especially neutralizing antibodies, have been shown to be associated with a lower incidence ofC. difficileinfection (CDI) recurrence, and antibody levels are predictive of asymptomatic colonization. The development of an assay to detect the presence of neutralizing antibodies in animal and human sera for the evaluation of vaccine efficacy is highly desired. We have developed such an assay, which allows for the quantification of the effect of toxins on eukaryotic cells in an automated manner. We describe here the optimization of this assay to measure toxin potency as well as neutralizing antibody (NAb) activity againstC. difficiletoxins using a design-of-experiment (DOE) methodology. Toxin concentration and source, cell seeding density, and serum-toxin preincubation time were optimized in the assay using Vero cells. The assay was shown to be robust and to produce linear results across a range of antibody concentrations. It can be used to quantify neutralizing antibodies in sera of monkeys and hamsters immunized withC. difficiletoxoid vaccines. This assay was shown to correlate strongly with traditional assays which rely on labor-intensive methods of determining neutralizing antibody titers by visual microscopic inspection of intoxicated-cell monolayers. This assay has utility for the selection and optimization ofC. difficilevaccine candidates.

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