Development and standardization of an in-house indirect ELISA for detection of duck antibody to fowl cholera

Poolperm, Pichayanut; Varinrak, Thanya; Kataoka, Yuki; Tragoolpua, Khajornsak; Sawada, Takuo; Sthitmatee, Nattawooti

doi:10.1016/j.mimet.2017.08.018

Cited by 15 publications

(13 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Bayesian latent class analysis is a statistical procedure for estimating the sensitivity and specificity of a diagnostic test in the absence of a gold standard assay. A Bayesian model has been used to estimate the sensitivity and specificity of various diagnostic techniques [27][28][29][30][31]. In this study, a Bayesian model was performed to estimate the sensitivity and specificity of an in-house indirect ELISA with two HRP-conjugated antibodies, and these values were compared to those of the HI test.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, the accuracy of newly developed diagnostic tests should be estimated appropriately in the absence of a gold standard. Recently, the Bayesian approach has been successfully applied to estimate the degrees of sensitivity and specificity in the absence of a gold standard [27,28,30,31]. The advantage of having access to Bayesian statistical inferences is the ability to simply model all of the uncertainties of the parameters with regard to probability [27].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

et al. 2020

Self Cite

View full text Add to dashboard Cite

Background: Feline panleukopenia virus (FPV) is an etiologic pathogen of feline panleukopenia that infects all members of Felidae including tigers (Panthera tigris). Vaccinations against FPV among wild felid species have long been practiced in zoos worldwide. However, few studies have assessed the tiger immune response post-vaccination due to the absence of a serological diagnostic tool. To address these limitations, this study aimed to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) for the monitoring of tiger antibody levels against the feline panleukopenia vaccine by employing the synthesized subunit capsid protein VP2. An in-house horseradish peroxidase (HRP) conjugated rabbit anti-tiger immunoglobulin G (IgG) polyclonal antibody (HRP-anti-tiger IgG) was produced in this study and employed in the assay. It was then compared to a commercial HRP-conjugated goat anti-cat IgG (HRPanti-cat IgG). Sensitivity and specificity were evaluated using the Bayesian model with preferential conditional dependence between HRP-conjugated antibody-based ELISAs and hemagglutination-inhibition (HI) tests. Results: The posterior estimates for sensitivity and specificity of two indirect ELISA HRP-conjugated antibodies were higher than those of the HI test. The sensitivity and specificity of the indirect ELISA for HRP-anti-tiger IgG and HRP-anticat IgG were 86.5, 57.2 and 86.7%, 64.6%, respectively, while the results of the HI test were 79.1 and 54.1%. In applications, 89.6% (198/221) and 89.1% (197/221) of the tiger serum samples were determined to be seropositive by indirect ELISA testing against HRP-anti-tiger and HRP-anti-cat, respectively.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

“…Bayesian latent class analysis is a statistical procedure for estimating the sensitivity and specificity of a diagnostic test in the absence of a gold standard assay. A Bayesian model has been used to estimate the sensitivity and specificity of various diagnostic techniques [27][28][29][30][31]. In this study, a Bayesian model was performed to estimate the sensitivity and specificity of an in-house indirect ELISA with two HRPconjugated antibodies, and these values were compared to the HI test.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

Areewong

Rittipornlertrak

Nambooppha

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Background: Feline panleukopenia virus (FPV) is an etiologic pathogen of feline panleukopenia that infects all members of Felidae including tigers (Panthera tigris). Vaccination against FPV among wild felid species has long been practiced in zoos worldwide. However, few studies have assessed tiger immune response post-vaccination due to the absence of a serological diagnostic tool. To address these limitations, this study aimed to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) for the monitoring of tiger antibody levels against the feline panleukopenia vaccine by employing the synthesized subunit capsid protein VP2. An in-house horseradish peroxidase (HRP) conjugated rabbit anti-tiger immunoglobulin G (IgG) polyclonal antibody (HRP-anti-tiger IgG) was produced in this study and employed in the assay. It was then compared to a commercial HRP-conjugated goat anti-cat IgG (HRP-anti-cat IgG). Sensitivity and specificity were evaluated using the Bayesian model with conditional dependence being assumed between both HRP-conjugated antibody-based ELISAs and hemagglutination-inhibition (HI) tests. Results: The posterior estimates for sensitivity and specificity of two indirect ELISA HRP-conjugated antibodies were higher than those of the HI test. The sensitivity and specificity of indirect ELISA for HRP-anti-tiger IgG and HRP-anti-cat IgG were 86.5%, 57.2% and 86.7%, 64.6%, respectively, while the results of the HI test were 79.1 and 54.1%. In applications, 89.6% (198/221) of tiger serum samples were determined to be seropositive by indirect ELISA testing. Conclusion: The results support evidence that an in-house indirect ELISA developed in this study could be used as a serological tool for the effective detection of tiger antibodies.

show abstract

“…To improve the sensitivity of the ic ELISA, we optimized different parameters, including the antibody-working concentration, coating concentration, coating time, blocking solution, diluted concentration of GAMIgG-HRP, and time of TMB colorization. Concentrations of the working antibody and coating antigen were optimized by checkerboard titration using DES-MCPE-OVA as the coating antigen 25 . Briefly, the DES-MCPE-OVA was coated on the ELISA plate at concentrations of 0.1, 0.2, 0.5, 1.0, 2.0, and 4.0 μg/ mL and the mAbs towards DES were diluted with PBS at dilutions of 1:4.0 × 10 3 , 1:8.0 × 10 3 , 1:1.6 × 10 4 , 1:3.2 × 10 4 , 1:6.4 × 10 4 , 1:1.28 × 10 5 , 1:1.28 × 10 5 , and 1:5.12 × 10 5 .…”

Section: Optimization Of the Ic Elisamentioning

confidence: 99%

Hapten Synthesis and the Development of an Ultrasensitive Indirect Competitive ELISA for the Determination of Diethylstilbestrol in Food Samples

Yang

Wang

Song

et al. 2020

Sci Rep

View full text Add to dashboard Cite

An ultrasensitive indirect competitive enzyme-linked immunosorbent assay (ic ELISA) using monoclonal antibodies (mAbs) was developed for the specific detection of diethylstilbestrol (DES) residues. To establish an ELISA based on mAbs, hapten diethylstilbestrol mono-carboxypropyl-ether (DES-MCPE) was chemically synthetized and then conjugated to bovine serum albumin (BSA) for immunization in mice. This ic ELISA was further optimized for DES determination. The sensitivity of the ic ELISA was found to be 0.49 μg/kg and the limit of detection was 0.075 μg/kg. DES residues in salmon meat and pork were tested with the recovery range from 74.0 to 85.2% and the coefficient of variation (CV) was less than 10%. Parallel analysis of DES samples from salmon meat showed comparable results from the ic ELISA with high-performance liquid chromatography. The ic ELISA provides a useful screening method for the quantitative detection of DES residues in animal-derived food.

show abstract

Development and standardization of an in-house indirect ELISA for detection of duck antibody to fowl cholera

Cited by 15 publications

References 20 publications

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

Evaluation of an in-house indirect enzyme-linked immunosorbent assay of feline panleukopenia VP2 subunit antigen in comparison to hemagglutination inhibition assay to monitor tiger antibody levels by Bayesian approach

Hapten Synthesis and the Development of an Ultrasensitive Indirect Competitive ELISA for the Determination of Diethylstilbestrol in Food Samples

Contact Info

Product

Resources

About