scholarly journals The Dengue ED3 Dot Assay, a Novel Serological Test for the Detection of Denguevirus Type-Specific Antibodies and Its Application in a Retrospective Seroprevalence Study

Viruses ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 304 ◽  
Author(s):  
Heidi Auerswald ◽  
Leonard Klepsch ◽  
Sebastian Schreiber ◽  
Janne Hülsemann ◽  
Kati Franzke ◽  
...  
Keyword(s):  
Specific Antibody ◽  
Serological Test ◽  
Cohort Analysis ◽  
Neutralizing Antibody ◽  
Elisa Test ◽  
Antibody Responses ◽  
Protein Domain ◽  
Antibody Detection ◽  
Serum Samples ◽  
Domain 3

: There are four distinct antigenic serotypes of dengue viruses (DENV-1-4). Sequential infections with different serotypes lead to crossreactive but also serotypespecific neutralizing antibody responses. Neutralization assays are considered as gold standard for serotype-specific antibody detection. However, for retrospective seroprevalence studies, access to large serum quantities is limited making neutralization assays well-nigh impossible. Therefore, a serological test, wasting only 10 µL serum, was developed using fusion proteins of maltose binding protein and E protein domain 3 (MBP-ED3) as antigens. Twelve MBP-ED3 antigens for DENV-1-4, three MBP-ED3 antigens for WNV, JEV, and TBEV, and MBP were dotted onto a single nitrocellulose strip. ED3 dot assay results were compared to virus neutralization and ED3 ELISA test results, showing a >90% accordance for DENV-1 and a 100% accordance for DENV-2, making the test specifically useful for DENV-1/-2 serotype-specific antibody detection. Since 2010, DENV-1 has replaced DENV-2 as the dominant serotype in Cambodia. In a retrospective cohort analysis, sera collected during the DENV-1/-2 endemic period showed a shift to DENV-2-specific antibody responses in 2012 paralleled by the decline of DENV-2 infections. Altogether, the ED3 dot assay is a serum-, time- and money-saving diagnostic tool for serotype-specific antibody detection, especially when serum samples are limited.

Experimental Schistosoma bovis infection in goats. Circulating antigen and antibody responses to egg and adult worm antigens during infection and following treatment with praziquantel

Parasitology ◽  
1996 ◽  
Vol 113 (4) ◽  
pp. 367-375 ◽  
Author(s):  
M. V. Johansen ◽  
Y. Fillié ◽  
J. Monrad ◽  
N. Ø. Christensen ◽  
A. Deelder
Keyword(s):  
Adult Worm ◽  
Specific Antibody ◽  
Post Treatment ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Faecal Samples ◽  
Schistosoma Bovis ◽  
Worm Recovery ◽  
Circulating Antigen ◽  
Positive Correlations

SUMMARYCirculating antigen levels and antibody responses in Schistosoma bovis-infected West African Dwarf goats were evaluated during infection and following treatment with praziquantel (60 mg/kg) 13 weeks post-infection. One day, 1 week and 4 weeks post-treatment, subgroups of goats were sacrificed and perfused for worm recovery. For comparison, parasite-free control animals were included. Blood and faecal samples were collected biweekly. Two gut-associated schistosome antigens, circulating cathodic and circulating anodic antigen (CCA and CAA) and 3 specific antibody responses (total Ig, IgG and IgM) were measured. For specific antibody detection, crude S. bovis adult worm and egg homogenates were used. The level of CCA in the infected groups was significantly elevated from the time of onset of egg excretion onwards. However, following treatment, the CCA litres dropped to control levels within 1 week post-treatment. Strong positive correlations were found between CCA levels and worm counts and faecal egg counts during peak egg excretion. The correlations of CAA and specific antibody litres to egg and worm counts were poor. The antibody responses were all significantly elevated in the infected goats during patency, but only marginally affected by the treatment. Hence, CCA proved to be superior by correlating strongly to the level of infection and by being a sensitive indicator of the effect of treatment.

scholarly journals Systemic and local antibodies induced by an experimental inactivated vaccine against bovine herpesvirus type 1

2011 ◽  
Vol 41 (2) ◽  
pp. 307-313
Author(s):  
Maria do Carmo Cilento ◽  
Edviges Maristela Pituco ◽  
Ricardo Spacagna Jordão ◽  
Cláudia Pestana Ribeiro ◽  
Moacir Marchiori Filho ◽  
...  
Keyword(s):  
Bovine Herpesvirus ◽  
Neutralization Assay ◽  
Elisa Test ◽  
Antibody Responses ◽  
Inactivated Vaccine ◽  
Serum Samples ◽  
Post Vaccination ◽  
Herpesvirus Type ◽  
Virus Neutralization Assay ◽  
Igg2 Antibody

An experimental inactivated vaccine against bovine herpesvirus-1 (BoHV-1) was produced aiming to evaluate the systemic and local antibody responses in 12 seronegative heifers, after vaccination and revaccination. Serum samples were submitted to virus neutralization assay and to ELISA test for detection of IgG1 and IgG2 isotypes. Nasal secretion samples were submitted to the same ELISA test for detection of IgG1 and IgG2 isotypes. The results showed that moderate to high neutralizing titres and IgG1 and IgG2 antibody responses were induced after the second vaccination in the serum and in nasal secretions up to 114 days post vaccination. IgG2 antibodies were the prevalent isotype for most of the post-vaccination period. The results indicate that BoHV-1 experimental inactivated vaccine elicited potentially protective IgG1 and IgG2 antibody levels, both in the systemic and mucosal compartments.

scholarly journals Rapid Detection of Serum Antibody by Dual-Path Platform VetTB Assay in White-Tailed Deer Infected with Mycobacterium bovis

2013 ◽  
Vol 20 (6) ◽  
pp. 907-911 ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
Rena Greenwald ◽  
Javan Esfandiari ◽  
Daniel J. O'Brien ◽  
Stephen M. Schmitt ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Mycobacterium Bovis ◽  
Skin Testing ◽  
Serum Antibody ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Serum Samples ◽  
Content Type ◽  
In The Wild ◽  
Free Ranging

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.

scholarly journals Isotype-specific antibody responses to foot-and-mouth disease virus in sera and secretions of ‘carrier’ and ‘non-carrier’ cattle

1996 ◽  
Vol 117 (2) ◽  
pp. 349-360 ◽  
Author(s):  
J. S. Salt ◽  
G. Mulcahy ◽  
R. P. Kitching
Keyword(s):  
Disease Virus ◽  
Specific Antibody ◽  
Statistical Significance ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Antibody Responses ◽  
Upper Respiratory Tract ◽  
Serum Samples ◽  
Mouth Disease Virus ◽  
Foot And Mouth

SummaryIsotype-specific antibody responses to foot-and-mouth disease virus (FMDV) were measured in the sera and upper respiratory tract secretions of vaccinated and susceptible cattle challenged with FMDV by direct contact or by intranasal inoculation. A comparison was made between cattle that eliminated FMDV and those that developed and maintained a persistent infection. Serological and mucosal antibody responses were detected in all animals after challenge. IgA and 1gM were detected before the development of IgG1and IgG2responses. 1gM was not detected in vaccinated cattle. Challenge with FMDV elicited a prolonged biphasic secretory antibody response in FMDV ‘carrier’ animals only. The response was detected as FMDVspecific IgA in both mucosal secretions and serum samples, which gained statistical significance (P< 0·05) by 5 weeks after challenge. This observation could represent the basis of a test to differentiate vaccinated and/or recovered convalescent cattle from FMDV ‘carriers’.

scholarly journals Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

2017 ◽  
Vol 61 (2) ◽  
pp. 163-171 ◽  
Author(s):  
Wendy González ◽  
Luis G. Giménez-Lirola ◽  
Ashley Holmes ◽  
Sergio Lizano ◽  
Christa Goodell ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Serum Antibody ◽  
Population Monitoring ◽  
Actinobacillus Pleuropneumoniae ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Post Inoculation ◽  
Serum Samples ◽  
Experimental Conditions ◽  
And Control

AbstractIntroduction:The prevention and control ofActinobacillus pleuropneumoniaein commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety ofA. pleuropneumoniaeantigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique toA. pleuropneumoniaeand produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods:Four groups of pigs (six pigs per group) were inoculated withA. pleuropneumoniaeserovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results:All pigs inoculated withA. pleuropneumoniaeserovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion:This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring forA. pleuropneumoniae.

scholarly journals Expansion of SARS-CoV-2-specific Antibody-secreting Cells and Generation of Neutralizing Antibodies in Hospitalized COVID-19 Patients

2020 ◽  
Author(s):  
Renata Varnaitė ◽  
Marina García ◽  
Hedvig Glans ◽  
Kimia T. Maleki ◽  
John Tyler Sandberg ◽  
...  
Keyword(s):  
B Cell ◽  
Specific Antibody ◽  
Neutralizing Antibodies ◽  
Cell Activation ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
B Cell Activation ◽  
Immunological Parameters ◽  
Antibody Secreting Cell ◽  
Antibody Levels

AbstractCoronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged in late 2019 and has since become a global pandemic. Pathogen-specific antibodies are typically a major predictor of protective immunity, yet B cell and antibody responses during COVID-19 are not fully understood. Here, we analyzed antibody-secreting cell (ASC) and antibody responses in twenty hospitalized COVID-19 patients. The patients exhibited typical symptoms of COVID-19, and presented with reduced lymphocyte numbers and increased T cell and B cell activation. Importantly, we detected an expansion of SARS-CoV-2 nucleocapsid protein-specific ASCs in all twenty COVID-19 patients using a multicolor FluoroSpot assay. Out of the 20 patients, 16 had developed SARS-CoV-2-neutralizing antibodies by the time of inclusion in the study. SARS-CoV-2-specific IgA, IgG and IgM antibody levels positively correlated with SARS-CoV-2-neutralizing antibody titers, suggesting that SARS-CoV-2-specific antibody levels may reflect the titers of neutralizing antibodies in COVID-19 patients during the acute phase of infection. Lastly, we showed that interleukin 6 (IL-6) and C-reactive protein (CRP) concentrations were higher in serum of patients who were hospitalized for longer, supporting the recent observations that IL-6 and CRP could be used to predict COVID-19 severity. Altogether, this study constitutes a detailed description of clinical and immunological parameters in twenty COVID-19 patients, with a focus on B cell and antibody responses, and provides tools to study immune responses to SARS-CoV-2 infection and vaccination.

scholarly journals Rapid, Simple, Quantitative, and Highly Sensitive Antibody Detection for Lyme Disease

2010 ◽  
Vol 17 (6) ◽  
pp. 904-909 ◽  
Author(s):  
Peter D. Burbelo ◽  
Alexandra T. Issa ◽  
Kathryn H. Ching ◽  
Jeffrey I. Cohen ◽  
Michael J. Iadarola ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Dynamic Range ◽  
Independent Set ◽  
Antibody Responses ◽  
Peptide Sequence ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Characteristic Analysis ◽  
Serum Samples ◽  
Serological Tests

ABSTRACT There is currently a need for improved serological tests for the diagnosis and monitoring of Lyme disease, an infection caused by Borrelia burgdorferi. In the present study, we evaluated luciferase immunoprecipitation systems (LIPSs) for use for profiling of the antibody responses to a panel of B. burgdorferi proteins for the diagnosis of Lyme disease. Initially, serum samples from a cohort of patients and controls (n = 46) were used for training and were profiled by the use of 15 different B. burgdorferi antigen constructs. For the patient sera, the antibody responses to several B. burgdorferi antigens, including VlsE, flagellin (FlaB), BmpA, DbpA, and DbpB, indicated that the antigens had high levels of immunoreactivity. However, the best diagnostic performance was achieved with a synthetic protein, designated VOVO, consisting of a repeated antigenic peptide sequence, VlsE-OspC-VlsE-OspC, Analysis of an independent set of serum samples (n = 139) used for validation showed that the VOVO LIPS test had 98% sensitivity (95% confidence interval [CI], 93% to 100%; P < 0.0001) and 100% specificity (95% CI, 94% to 100%; P < 0.0001). Similarly, the C6 peptide enzyme-linked immunosorbent assay (ELISA) also had 98% sensitivity (95% CI, 93% to 100%; P < 0.0001) and 98% specificity (95% CI, 90% to 100%; P < 0.0001). Receiver operating characteristic analysis revealed that the rates of detection of Lyme disease by the LIPS test and the C6 ELISA were not statistically different. However, the VOVO LIPS test displayed a wide dynamic range of antibody detection spanning over 10,000-fold without the need for serum dilution. These results suggest that screening by the LIPS test with VOVO and other B. burgdorferi antigens offers an efficient quantitative approach for evaluation of the antibody responses in patients with Lyme disease.

scholarly journals Borrelia burgdorferi Spirochetes That Harbor Only a Portion of the lp28-1 Plasmid Elicit Antibody Responses Detectable with the C6 Test for Lyme Disease

2006 ◽  
Vol 14 (1) ◽  
pp. 90-93 ◽  
Author(s):  
Monica E. Embers ◽  
Gary P. Wormser ◽  
Ira Schwartz ◽  
Dale S. Martin ◽  
Mario T. Philipp
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Antibody Responses ◽  
Antibody Detection ◽  
23S Rrna ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Altered Expression ◽  
Genetic Region ◽  
Length Polymorphisms

ABSTRACT Detection of antibody to C6, a peptide that reproduces the sequence of the sixth invariable region within the central domain of the VlsE protein of Borrelia burgdorferi, is used currently for the serologic diagnosis of Lyme disease in humans. B. burgdorferi isolates taken from infected humans can be categorized into specific genetic subtypes (designated RST1, -2, and -3) by restriction fragment length polymorphisms in the 16S to 23S rRNA spacer sequence. Many of these, usually categorized as RST2, retain only segments of the linear plasmid lp28-1, which encodes VlsE. The VlsE genetic region is retained, but altered expression of this molecule could affect diagnosis by the C6 enzyme-linked immunosorbent assay (ELISA). Serum samples from patients infected with each of the three genotypes and from mice infected with three RST2 isolates were tested with the C6 ELISA. Such isolates elicited marked C6 responses in infected mice. The sensitivity of C6 antibody detection in patients infected with RST2 spirochetes was statistically indistinguishable from detection of RST1 and RST3 infections. These findings demonstrate that diagnosis by C6 ELISA remains effective for infection with all B. burgdorferi genotypes, including those with incomplete lp28-1 plasmids.

scholarly journals SARS-CoV-2 antibodies remain detectable 12 months after infection and antibody magnitude is associated with age and COVID-19 severity

2021 ◽  
Author(s):  
Eric D. Laing ◽  
Nusrat J. Epsi ◽  
Stephanie A. Richard ◽  
Emily C. Samuels ◽  
Wei Wang ◽  
...  
Keyword(s):  
Symptom Onset ◽  
Neutralizing Antibodies ◽  
Natural Infection ◽  
Geometric Mean ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Serum Samples ◽  
Military Hospitals ◽  
Igg Binding ◽  
One Year

ABSTRACTImportanceThe persistence of SARS-CoV-2 antibodies may be a predictive correlate of protection for both natural infections and vaccinations. Identifying predictors of robust antibody responses is important to evaluate the risk of re-infection / vaccine failure and may be translatable to vaccine effectiveness.ObjectiveTo 1) determine the durability of anti-SARS-CoV-2 IgG and neutralizing antibodies in subjects who experienced mild and moderate to severe COVID-19, and 2) to evaluate the correlation of age and IgG responses to both endemic human seasonal coronaviruses (HCoVs) and SARS-CoV-2 according to infection outcome.DesignLongitudinal serum samples were collected from PCR-confirmed SARS-CoV-2 positive participants (U.S. active duty service members, dependents and military retirees, including a range of ages and demographics) who sought medical treatment at seven U.S. military hospitals from March 2020 to March 2021 and enrolled in a prospective observational cohort study.ResultsWe observed SARS-CoV-2 seropositivity in 100% of inpatients followed for six months (58/58) to one year (8/8), while we observed seroreversion in 5% (9/192) of outpatients six to ten months after symptom onset, and 18% (2/11) of outpatients followed for one year. Both outpatient and inpatient anti-SARS-CoV-2 binding-IgG responses had a half-life (T1/2) of >1000 days post-symptom onset. The magnitude of neutralizing antibodies (geometric mean titer, inpatients: 378 [246-580, 95% CI] versus outpatients: 83 [59-116, 95% CI]) and durability (inpatients: 65 [43-98, 95% CI] versus outpatients: 33 [26-40, 95% CI]) were associated with COVID-19 severity. Older age was a positive correlate with both higher IgG binding and neutralizing antibody levels when controlling for COVID-19 hospitalization status. We found no significant relationships between HCoV antibody responses and COVID-19 clinical outcomes, or the development of SARS-CoV-2 neutralizing antibodies.Conclusions and RelevanceThis study demonstrates that humoral responses to SARS-CoV-2 infection are robust on longer time-scales, including those arising from milder infections.However, the magnitude and durability of the antibody response after natural infection was lower and more variable in younger participants who did not require hospitalization for COVID-19. These findings support vaccination against SARS-CoV-2 in all suitable populations including those individuals that have recovered from natural infection.

Sign in / Sign up

Export Citation Format

Share Document