Low levels of genetic diversity associated with evidence of negative selection on the Babesia bovis apical membrane antigen 1 from parasite populations in Thailand

Rittipornlertrak, Amarin; Nambooppha, Boondarika; Simking, P.; Punyapornwithaya, Veerasak; Tiwananthagorn, Saruda; Jittapalapong, Sathaporn; Chung, Yang-Tsung; Sthitmatee, Nattawooti

doi:10.1016/j.meegid.2017.08.009

Cited by 19 publications

(20 citation statements)

References 65 publications

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“…The VP2 subunit antigen was predicted based on the amino acid sequence accession number ABN70938.1 using the linear and conformational epitope prediction software as has been described in the previous report [ 37 ]. The selected amino acid sequence (WNPIQQMSINVDNQFNYVPNNIGAMKIVYEKSQLAPRKLY) was then further synthesized (Genscript, Piscataway, New Jersey, USA).…”

Section: Methodsmentioning

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

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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.

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Section: Methodsmentioning

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

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“…The VP2 subunit antigen was predicted based on the amino acid sequence accession number ABN70938.1 using the linear and conformational epitope prediction software as was described in the previous report [37]. The selected amino acid sequence (WNPIQQMSINVDNQFNYVPNNIGAMKIVYEKSQLAPRKLY) was then further synthesized (Genscript, Piscataway, New Jersey, USA).…”

Section: Capsid Protein or Viral Protein 2 (Vp2) Subunit Antigen Predmentioning

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

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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.

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“…A BbAMA-1 amino acid sequence retrieved from UniProtKB database (entry no. A7ASF6) was subjected to bioinformatics analysis to predict both linear and conformational B-cell epitope in the PAN motif of domain I as described by Rittipornlertrak et al (2017) [26]. The genetic diversity of the predicted epitope was visualized by multiply aligning the corresponding amino acid sequences from various B. bovis isolates, using the MUSCLE algorithm (http://www.ebi.ac.uk/Tools/msa/muscle/) [44].…”

Section: Prediction Analysis and Synthesis Of A B-cell Epitopementioning

confidence: 99%

“…However, the use of AMA-1 in malaria vaccine formulations is constrained due to its high genetic diversity [23][24]. In contrast, the B. bovis AMA-1 (BbAMA-1) is highly conserved in B. bovis [25][26], and therefore is a target for diagnostic assays [27]. The BbAMA-1 composes of three domains of ectodomain (domain I-III) [11].…”

Section: Introductionmentioning

confidence: 99%

Structural and immunological characterization of an epitope within the PAN motif of ectodomain I in Babesia bovis apical membrane antigen 1

Rittipornlertrak

Nambooppha

Muenthaisong

et al. 2020

Preprint

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Background Bovine babesiosis caused by Babesia bovis (B. bovis) affects the cattle industry worldwide with high mobility and mortality. Live-attenuated vaccines are currently used in some of the endemic countries, but their wide use is limited due to various reasons. Although recombinant vaccines have been proposed as an alternative to the live vaccines, such vaccines are not commercially available to date. Apical membrane antigen-1 (AMA-1) is one of the leading candidates for vaccine development against diseases caused by apicomplexan parasite species. In this study, we predicted an epitope from the plasminogen, apple and nematode (PAN) motif of domain I in the B. bovis AMA-1 (BbAMA-1) using a combination of linear and conformational B-cell epitope prediction software. The selected epitope was bioinformatically analyzed, synthesized as a peptide (sBbAMA-1), and then used to immunize a rabbit. Results The anti-sBbAMA-1 serum obtained was evaluated for its growth- and invasion-inhibitory effects on B. bovis merozoites in vitro. Our results demonstrated that the predicted BbAMA-1 epitope, which is located on surface-exposed α-helix of PAN motif in domain I at the apex area, elicits antibodies capable of recognizing the native BbAMA-1 in immunoassays. Importantly, as compared to the control groups, the rabbit anti-sBbAMA-1 serum at dilution of 1:5 significantly inhibited (p < 0.05) the growth of B. bovis merozoites by approximately 50–70% on day 3 and 4 of cultivation and the invasion of merozoites by approximately 60% within 4 h of incubation. Conclusion Our results indicate the epitope predicted from the PAN motif of BbAMA-1 domain I is neutralization-sensitive and may serve as a target antigen for vaccine development against bovine babesiosis caused by B. bovis.

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Low levels of genetic diversity associated with evidence of negative selection on the Babesia bovis apical membrane antigen 1 from parasite populations in Thailand

Cited by 19 publications

References 65 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

Structural and immunological characterization of an epitope within the PAN motif of ectodomain I in Babesia bovis apical membrane antigen 1

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