scholarly journals Improved Specificity of a Multiplex Immunoassay for Quantitation of Anti-Diphtheria Toxin Antibodies with the Use of Diphtheria Toxoid

2011 ◽  
Vol 18 (7) ◽  
pp. 1183-1186 ◽  
Author(s):  
Pieter G. M. van Gageldonk ◽  
Christina von Hunolstein ◽  
Fiona R. M. van der Klis ◽  
Guy A. M. Berbers

ABSTRACTA nonspecific binding of antibodies to diphtheria toxin, especially in adult serum samples, was observed in our diphtheria-tetanus-pertussis multiplex immunoassay (DTaP4 MIA). This can be significantly reduced by the use of diphtheria toxoid, achieving a good correlation with the Vero cell neutralization test and the toxin binding inhibition assay.

2017 ◽  
Vol 241 ◽  
pp. 15-23 ◽  
Author(s):  
Rutger M. Schepp ◽  
Guy A.M. Berbers ◽  
José A. Ferreira ◽  
Johan H. Reimerink ◽  
Fiona R. van der Klis

Author(s):  
P.M. Hale ◽  
M. Liebert ◽  
J.C. Sisson ◽  
T. L. Whiteside ◽  
NJ. Hopwood ◽  
...  

2021 ◽  
Author(s):  
Natalie E Hofmann ◽  
Marica Grossegesse ◽  
Markus Neumann ◽  
Lars Schaade ◽  
Andreas Nitsche

Background: High-throughput detection of neutralizing antibodies against SARS-CoV-2 presents a valuable tool for vaccine trials or investigations of population immunity. We evaluate the performance of the first commercial surrogate virus neutralization test (sVNT, GenScript Biotech) against SARS-CoV-2 plaque reduction neutralization test (PRNT) in convalescent and vaccinated individuals. We compare it to five other ELISAs, two of which are designed to detect neutralizing antibodies. Results: In 491 pre-vaccination serum samples, sVNT missed 23.6% of PRNT-positive samples when using the manufacturer-recommended cutoff of 30% binding inhibition. Introducing a equivocal area between 15 and 35% maximized sensitivity and specificity against PRNT to 72.8-93.1 % and 73.5-97.6%, respectively. The overall diagnostic performance of the other ELISAs for neutralizing antibodies was below that of sVNT. Vaccinated individuals exhibited higher antibody titers by PRNT (median 119.8, IQR 56.7-160) and binding inhibition by sVNT (median 95.7, IQR 88.1-96.8) than convalescent patients (median 49.1, IQR 20-62; median 52.9, IQR 31.2-76.2). Conclusion: GenScript sVNT is suitable to screen for SARS-CoV-2-neutralizing antibodies; however, to obtain accurate results, confirmatory testing by PRNT in a equivocal area is required. This equivocal area may require adaptation for use in vaccinated individuals, due to higher antibody titers.


1977 ◽  
Vol 145 (1) ◽  
pp. 187-203 ◽  
Author(s):  
R D Stout ◽  
D B Murphy ◽  
H O McDevitt ◽  
L A Herzenberg

Treatment of splenic T lymphocytes with anti-Ia antiserum inhibits the binding of antigen-antibody (AgAb) complexes to the majority (less than 50%) of Fc receptor-positive (FcR+) T cells. A similar inhibition was observed with anti-H-2D and anti-H-2K sera but not with anti-Thy 1.2. Despite the presence of Ia determinants on peripheral T cells, as established by the inhibition of AgAb binding, Ia could not be detected on peripheral T cells by immunofluorescence assays. Data obtained with the AgAb-binding inhibition assay indicate that determinants controlled by loci mapping in the I-A and I-C, S, or G regions are present on the FcR+ T cells. Evidence is presented that subpopulations of T cells within the FcR+ T-cell population may be distinguishable on the basis of which I-region-controlled determinant is expressed. The data are discussed in terms of phenotypic and functional heterogeneity of T lymphocytes.


2000 ◽  
Vol 125 (1) ◽  
pp. 105-112 ◽  
Author(s):  
M. BERGAMINI ◽  
P. FABRIZI ◽  
S. PAGANI ◽  
A. GRILLI ◽  
R. SEVERINI ◽  
...  

This study evaluated whether a correlation exists between carriage of corynebacteria and the lack of immunity to diphtheria toxoid. Samples of both nasal and pharyngeal secretions were taken from 500 apparently healthy subjects of both sexes and of all ages and inoculated onto Tinsdale's medium. A serum sample was also taken for ELISA test to determine the titre of diphtheria toxin antibodies. None of the subjects carried Corynebacterium diphtheriae. Ninety-three strains of Corynebacterium spp. were isolated from 93 subjects and 86 of these were classified to species or group level by biochemical tests. C. xerosis was the most common (25·8%) followed by C. pseudodiphthericum (16·1%), C. jeikeium and C. striatum (both 10·8%), and C. urealyticum (9·7%). Three other species accounted for approximately 20% of strains and seven were unclassified as biochemically atypical corynebacteria. Non-protective antibodies to diphtheria toxin were found in 80 of the 93 subjects and a strong statistical association was demonstrated between carriage of corynebacteria and non-protective levels of anti-toxin antibodies. The remaining 13 subjects had protective levels of antitoxin antibodies. In contrast, only 45 of the 407 non-colonized subjects had non-protective antitoxin titres. The prevalence of carriage increased with age among males as did the percentage of non-protected subjects. The prevalence of female carriers of corynebacteria was significantly lower. Serum samples from 12 subjects with different antibody titres to diphtheria toxoid reacted to varying degrees with whole-cell lysates of a number of species of corynebacteria. The results suggest that a causal relationship may exist between nasopharyngeal carriage of corynebacteria and a low anti-diphtheria toxin immune response.


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