Development of an indirect ELISA to specifically detect antibodies against African swine fever virus: bioinformatics approaches

Gao, Zhan; Shao, Jing; Zhang, Guanglei; Ge, Sudan; Chang, Yung-Fu; Xiao, Lei; Chang, Huiyun

doi:10.1186/s12985-021-01568-2

Cited by 13 publications

(8 citation statements)

References 31 publications

(6 reference statements)

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“…These 82 serum samples were then analyzed using the optimized CD2v-iELISA method. The cut-off value was defined as the mean OD 450 value + 3 × the standard deviation (SD), and samples above this cut-off value were considered to be ASFV + ( 19 ).…”

Section: Methodsmentioning

confidence: 99%

Development of an indirect ELISA for the identification of African swine fever virus wild-type strains and CD2v-deleted strains

Jiang

et al. 2022

Front. Vet. Sci.

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African swine fever (ASF) is a potent infectious disease with detrimental effects on the global swine industry and no currently vaccine available. The emergence of low-virulence CD2v-deleted mutants manifested as non-hemadsorption (non-HAD) strains represents a significant challenge to the prevention and control of ASF. In this study, we aimed to establish an indirect ELISA (IELISA) method for the identification of ASFV wild-type and CD2v-deleted strains. We integrated the CD2v protein extracellular domain sequence (CD2v-Ex, 1–588 bp) of the highly pathogenic strain China/2018/AnhuiXCGQ into the genome of suspension culture-adapted Chinese hamster Ovary-S (CHO-S) cells using lentivirus vectors (LVs). By screening, we identified a monoclonal CHO-S cell line that stably expressed secretory CD2v-Ex Protein. We then used the purified CD2v-Ex Protein as the detection antigen to establish an indirect ELISA method (CD2v-IELISA) for identification of the ASFV wild-type and CD2v-Deleted (CD2v−) strains. The CD2v-IELISA method showed excellent specificity with no cross-reaction with serum samples infected with ASFV (CD2v−), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), porcine circovirus (PCV), porcine pseudorabies virus (PRV), swine foot and mouth disease virus (FMDV) and porcine epidemic diarrhea virus (PEDV). Furthermore, this method showed high sensitivity, allowing identification of ASFV-infected clinical serum samples up to a dilution of 1:2,560. The coefficient of variation both in and between batches was <10% with good reproducibility and a high compliance rate of 99.4%. This CD2v-IELISA method developed here is of great significance for the prevention, control and purification of ASFV.

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Section: Methodsmentioning

confidence: 99%

Development of an indirect ELISA for the identification of African swine fever virus wild-type strains and CD2v-deleted strains

Jiang

et al. 2022

Front. Vet. Sci.

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“…coli cells ELISA 88 hepatitis-positive serum samples/376 hepatitis C-negative serum samples/18 serum samples from non-hepatitis C diseases Sensitivity: 100% Specificity: 99.73% [ 106 ]/Brazil Human cytomegalovirus/cytomegalovirus rMEHCMV E. coli BL21(DE3) cells ELISA 12 cytomegalovirus-positive serum samples/1 serum sample from healthy individual rMEHCMV was recognized by human positive samples [ 107 ]/Brazil Canine coronavirus/SARS-CoV-2 rSP E. coli BL21(DE3) cells Indirect ELISA 64 coronavirus-positive serum samples/10 healthy dog serum samples 82.81% positive rate [ 108 ]/China Human coronavirus/SARS-CoV-2 Dx-SARS2-RBD and Dx-SARS2-noRBD E. coli BL21(DE3) cells ELISA 185 coronavirus-positive serum samples/256 serum samples from healthy individuals/94 serum samples from non-coronovirus diseases Dx-SARS2-RBD Sensitivity: 100% Specificity: 99.51-100% Dx-SARS2-noRBD Sensitivity: 100% Specificity: 99.21-100% [ 109 ]/Brazil African swine fever/African swine fever virus reMeP72 E. coli BL21(DE3) cells Colloidal gold-based immunochromatographic assay 139 swine serum samples Sensitivity: 85.7% Specificity: 97.6% [ 110 ]/China African swine fever/African swine fever virus m35 E . coli BL21 cells Indirect ELISA 78 positive serum samples/215 negative serum samples Sensitivity: 98.72% Specificity: 98.14% [ 111 ]/China Human mayaro fever/mayar...…”

Section: Rmps Applied In Disease Diagnosismentioning

confidence: 99%

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

Gonçalves,

Ribeiro,

Resende

et al. 2024

Microb Cell Fact

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Recombinant multiepitope proteins (RMPs) are a promising alternative for application in diagnostic tests and, given their wide application in the most diverse diseases, this review article aims to survey the use of these antigens for diagnosis, as well as discuss the main points surrounding these antigens. RMPs usually consisting of linear, immunodominant, and phylogenetically conserved epitopes, has been applied in the experimental diagnosis of various human and animal diseases, such as leishmaniasis, brucellosis, cysticercosis, Chagas disease, hepatitis, leptospirosis, leprosy, filariasis, schistosomiasis, dengue, and COVID-19. The synthetic genes for these epitopes are joined to code a single RMP, either with spacers or fused, with different biochemical properties. The epitopes’ high density within the RMPs contributes to a high degree of sensitivity and specificity. The RMPs can also sidestep the need for multiple peptide synthesis or multiple recombinant proteins, reducing costs and enhancing the standardization conditions for immunoassays. Methods such as bioinformatics and circular dichroism have been widely applied in the development of new RMPs, helping to guide their construction and better understand their structure. Several RMPs have been expressed, mainly using the Escherichia coli expression system, highlighting the importance of these cells in the biotechnological field. In fact, technological advances in this area, offering a wide range of different strains to be used, make these cells the most widely used expression platform. RMPs have been experimentally used to diagnose a broad range of illnesses in the laboratory, suggesting they could also be useful for accurate diagnoses commercially. On this point, the RMP method offers a tempting substitute for the production of promising antigens used to assemble commercial diagnostic kits.

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“…Bioinformatic methods using a machine learning approach serve as an effective strategy to identify vaccine candidates for human (Guo et al 2022;Hajialibeigi et al 2021; Kibria et al 2022) and swine pathogens, including ASFV (Gao et al 2021), in uenza A virus (Baratelli et al 2020;Fan et al 2018), and porcine circovirus type 2 (Bandrick et al 2020). Machine learning methods used to identify epitopes in the design of multi-epitope vaccines consider the biological presentation and activation of ASFV epitopes.…”

Section: Introductionmentioning

confidence: 99%

Multi-epitope vaccine design of African swine fever virus considering T cell and B cell immunogenicity

Chen,

Ho,

et al. 2024

Preprint

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T and B cell activation are equally important in triggering and orchestrating adaptive host responses to design multi-epitope African swine fever virus (ASFV) vaccines. However, few design methods have considered the trade-off between T and B cell immunogenicity when identifying promising ASFV epitopes. This work proposed a novel Pareto front-based ASFV screening method PFAS to identify promising epitopes for designing multi-epitope vaccines utilizing five ASFV Georgia 2007/1 sequences. To accurately predict T cell immunogenicity, four scoring methods were used to estimate the T cell activation in the four stages, including proteasomal cleavage probability, transporter associated with antigen processing transport efficiency, class I binding affinity of the major histocompatibility complex, and CD8 + cytotoxic T cell immunogenicity. PFAS ranked promising epitopes using a Pareto front method considering T and B cell immunogenicity. The coefficient of determination between the Pareto ranks of multi-epitope vaccines and survival days of swine vaccinations was R2 = 0.95. Consequently, PFAS scored complete epitope profiles and identified 72 promising top-ranked epitopes, including 46 CD2v epitopes, two p30 epitopes, 10 p72 epitopes, and 14 pp220 epitopes. PFAS is the first method of using the Pareto front approach to identify promising epitopes that considers the objectives of maximizing both T and B cell immunogenicity. The top-ranked promising epitopes can be cost-effectively validated in vitro. The Pareto front approach can be adaptively applied to various epitope predictors for bacterial, viral and cancer vaccine developments. The MATLAB code of the Pareto front method was available at https://github.com/NYCU-ICLAB/PFAS.

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Development of an indirect ELISA to specifically detect antibodies against African swine fever virus: bioinformatics approaches

Cited by 13 publications

References 31 publications

Development of an indirect ELISA for the identification of African swine fever virus wild-type strains and CD2v-deleted strains

Development of an indirect ELISA for the identification of African swine fever virus wild-type strains and CD2v-deleted strains

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

Multi-epitope vaccine design of African swine fever virus considering T cell and B cell immunogenicity

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