Single dilution Avidity-Blocking ELISA as an alternative to the Bovine Viral Diarrhea Virus neutralization test

Mahecha, O.L. Franco; Castells, Matías; Combessies, G.; Lavoria, María Ángeles; Wilda, Maximiliano; Mansilla, Florencia Celeste; Seki, Cristina; Grigera, Pablo R.; Capozzo, Alejandra Victoria

doi:10.1016/j.jviromet.2011.05.022

Cited by 18 publications

(11 citation statements)

References 32 publications

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“…In vitro neutralizing activity usually correlates with avidity (Bachmann et al 1997) which can be easily measured by ELISA. Correlation between avidity and VNT has been reported for vesicular stomatitis virus (Bachmann et al 1997), measles virus (Saika et al 2008), tick-borne encephalitis (Leonova & Pavlenko 2009) and bovine viral diarrhoea virus (Franco Mahecha et al 2011). We have also shown that the avidity indexes of anti-FMDV bovine antibodies are concordant with the VNT results (Lavoria et al 2012;).…”

Section: Introductionsupporting

confidence: 76%

“…These assays avoid the need of developing coating and detector antibodies for each particular strain and allow high-throughput assessment as they are performed with a single dilution of the sera, for detecting both total (IE) and high-avidity antibodies (AE). AEs are usually used to discriminate between chronic and acute infections, or as an alternative to neutralization assays, among other applications (Thomas & Morgan-Capner 1988;Enders & Knotek 1989;Schubert et al 1998;Fox et al 2006;Hamkar et al 2006;Franco Mahecha et al 2011). This strategy has been applied before to detach low binders and increase the specificity of the ELISA applied for FMDV diagnosis in cross-protection trials .…”

Section: Discussionmentioning

confidence: 99%

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Alternatives for the serological assessment of foot-and-mouth disease vaccine immunity in buffaloes (Bubalus bubalis)

Sala

Trotta

Mansilla

et al. 2017

Journal of Applied Animal Research

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Buffaloes are compulsory vaccinated against foot-and-mouth disease virus (FMDV) in many countries as part of the official control programmes. Serological testing aimed to indirectly assess herd immunity is currently performed using the same Enzyme-linked immunosorbent assays (ELISAs) applied for bovine sera, assuming an agreement between the ELISA's diagnostic results and those obtained using the virus neutralization test (VNT). Here we evaluated the accuracy of different ELISA tests to assess vaccine-induced antibodies against FMDV in buffalo's sera classified according to their VNT titres. Currently used liquid-phase blocking ELISA yielded very low specificity, producing high titres for many samples with low VNT titres. To increase specificity, we developed an indirect ELISA using purified 140S viral particles and an avidity single-dilution ELISA, which includes a urea washing step after the incubation of the diluted serum sample, to detach weak binders. Combining these two highthroughput single-dilution tests, an excellent concordance with VNT was achieved. This is the first study analysing the diagnostic agreement of traditional and novel serological tests with VNT for the indirect assessment of antibodies against FMDV capsid proteins in buffalo serum samples. ARTICLE HISTORY

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Section: Introductionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Alternatives for the serological assessment of foot-and-mouth disease vaccine immunity in buffaloes (Bubalus bubalis)

Sala

Trotta

Mansilla

et al. 2017

Journal of Applied Animal Research

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“…[11,12] But it was lower than an avidity blocking ELISA using drosophila expressed E2 antigen that showed a sensitivity of 98.10%. [32] This may be explained by the reason that most ELISAs probably detect preferentially IgG isotypes, while VNT detects both IgM and IgG isotypes. [33] A slightly lower specificity of the developed E2 ELISA was observed in our study when compared to the VNT.…”

Section: Discussionmentioning

confidence: 99%

Expression of Bovine Viral Diarrhea Virus Envelope Glycoprotein E2 in YeastPichia pastorisand its Application to an ELISA for Detection of BVDV Neutralizing Antibodies in Cattle

Behera

Mishra

Nema

et al. 2015

Journal of Immunoassay and Immunochemistry

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The aim of this article is to express envelope glycoprotein E2 of bovine viral diarrhea virus (BVDV) in yeast Pichia pastoris and its utility as a diagnostic antigen in ELISA. The BVDV E2 gene was cloned into the pPICZαA vector followed by integration into the Pichia pastoris strain X-33 genome for methanol-induced expression. SDS-PAGE and Western blot results showed that the recombinant BVDV E2 protein (72 kDa) was expressed and secreted into the medium at a concentration of 40 mg/L of culture under optimized conditions. An indirect ELISA was then developed by using the yeast-expressed E2 protein. Preliminary testing of 300 field cattle serum samples showed that the E2 ELISA showed a sensitivity of 91.07% and a specificity of 92.02% compared to the reference virus neutralization test. The concordance between the E2 ELISA and VNT was 91.67%. This study demonstrates feasibility of BVDV E2 protein expression in yeast Pichia pastoris for the first time and its efficacy as an antigen in ELISA for detecting BVDV neutralizing antibodies in cattle.

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“…The avidity of those antibodies was determined by incubating plates with 100 μL per well of 6 mol/L urea–PBS for 20 minutes after the first washing step . The ELISA then followed the protocol previously described.…”

Section: Methodsmentioning

confidence: 99%

Fusion of foreign T‐cell epitopes and addition of TLR agonists enhance immunity against Neospora caninum profilin in cattle

Mansilla¹,

Quintana

Cardoso

et al. 2016

Parasite Immunology

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We demonstrated recently that immunization with recombinant Neospora caninum profilin (rNcPRO) induces limited protection and a regulatory T-cell response in mice. The aim of this study was to evaluate the immune response elicited by rNcPRO in cattle and assess a strategy to enhance its immunogenicity, combining the addition of T-cell epitopes and immune modulators. We developed a chimeric recombinant profilin fused to functional T-cell epitopes present in the N-terminal sequence of vesicular stomatitis virus (VSV) glycoprotein G (rNcPRO/G). Groups of three cattle were immunized with two doses (2 weeks apart) of rNcPRO or rNcPRO/G formulated with alum hydroxide or a nanoparticulated soya-based adjuvant enriched with Toll-like receptor (TLR) 2 and TLR9 agonists, aimed to tackle the MyD88 pathway (AVECplus). rNcPRO induced only a primary immune response (IgM mediated), while antibodies in rNcPRO/G-vaccinated animals switched to IgG1 after the booster. The vaccine formulated with rNcPRO/G and AVECplus improved the production of systemic IFN-γ and induced long-term recall B-cell responses. Overall, our study provides data supporting the use of T-cell epitopes from VSV glycoprotein G and TLR agonists to enhance and modulate immunity to peptide antigens in bovines, particularly when using small proteins from parasites for which immune responses are usually feeble.

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Single dilution Avidity-Blocking ELISA as an alternative to the Bovine Viral Diarrhea Virus neutralization test

Cited by 18 publications

References 32 publications

Alternatives for the serological assessment of foot-and-mouth disease vaccine immunity in buffaloes (Bubalus bubalis)

Alternatives for the serological assessment of foot-and-mouth disease vaccine immunity in buffaloes (Bubalus bubalis)

Expression of Bovine Viral Diarrhea Virus Envelope Glycoprotein E2 in YeastPichia pastorisand its Application to an ELISA for Detection of BVDV Neutralizing Antibodies in Cattle

Fusion of foreign T‐cell epitopes and addition of TLR agonists enhance immunity against Neospora caninum profilin in cattle

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