Development and evaluation of a monoclonal antibody-based blocking ELISA to detect antibodies against the E2 protein of bovine viral diarrhea virus-1

Liu, Xinhuan; Cheng, Zilong; Zhang, Wenwen; Mao, Li; Pan, Zihao; Yang, Leilei; Liu, Maojun; Long, Yunfeng; Bai, Juan; Li, Wenliang

doi:10.1016/j.jviromet.2023.114851

Cited by 3 publications

(1 citation statement)

References 37 publications

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“…In recent times, BVDV infection and associated diseases have become widespread in China, leading to significant economic losses in the animal husbandry sector (Liu et al 2024). Up to now, there are mainly BVDV-1 species in China, while the BVDV-2 species are rarely found (Yang et al 2022).…”

Section: Discussionmentioning

confidence: 99%

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

2024

Int J Vet Sci

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Bovine viral diarrhea virus (BVDV) is an economically consequential animal pathogen that engenders substantial financial losses in the realm of cattle farming, and it maintains a prevalent presence on a global scale. Presently, the prevailing modality employed for its detection is enzyme-linked immunosorbent assay (ELISA), which necessitates the use of sophisticated instrumentation, protracts the duration of analysis, and is primarily suited for laboratory-based diagnostics. Regrettably, ELISAs do not lend themselves to convenient on-site detection within cattle farms. To surmount this predicament, colloidal gold particles were synthesized using tri-sodium citrate reduction, and subsequently harnessed to label the E2 protein (E2-Au). By amalgamating the gold standard binding release pad with colloidal E2-Au, we have devised a comprehensive assay system. The nitrocellulose membrane serves as the platform, whereby the detection line is adorned with E2-Au, while the quality control line is coated with proprietary polyclonal rabbit antibodies specific to E2. The method yields results within a span of 10-15 minutes, exhibiting an impressive total compliance rate of 93.36% (239/256) when compared to the commercial kit. The E2 test strip demonstrates specific reactivity with the anti-BVDV antibody, while avoiding any cross-reactivity with antibodies targeting Brucella, bovine foot and mouth disease, Mycobacterium bovis, bovine para-tuberculosis, bovine pasteurellosis, and bovine infectious rhinotracheitis. Consequently, the E2 strips offer heightened specificity, cost-effectiveness, and ease of operation, rendering them more convenient and expedient in comparison to ELISA kits. Utilizing the E2 strips, a comprehensive assessment encompassing 36 cattle farms and 2035 cattle in the vicinity of Xinjiang, China, was conducted. The positive rate within the group reached an impressive 91.67%, with individual positive rates standing at 54.05%.

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Section: Discussionmentioning

confidence: 99%

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

2024

Int J Vet Sci

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Development of a pan-genotypic monoclonal antibody-based competitive ELISA for the detection of antibodies against Bovine viral diarrhea virus

Qi,

Wang,

et al. 2024

Front. Immunol.

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IntroductionBovine viral diarrhea virus (BVDV), a positive-sense single-stranded RNA virus, causes significant economic losses in the cattle industry. Current diagnostic methods for BVDV exhibit variable sensitivity and specificity, underscoring the need for more rapid and accurate detection approaches. Here, we developed a novel competitive ELISA (cELISA) to detect antibodies against the BVDV E2 protein.Methods and resultsWe generated three monoclonal antibodies (mAbs)—3E6, 2D5, and 5B9—by immunizing mice with purified BVDV E2 protein expressed in Expi293F cells. Among these, mAb 3E6 displayed superior competitive binding abilities to the E2 protein, enabling effective differentiation between BVDV positive and negative sera. Remarkably, mAb 3E6 exhibited pan-genotypic recognition of various BVDV strains, including BVDV-1a, -1b, -1c, -1m, -1p, -1v, and -2a, while showing no cross-reactivity with the classical swine fever virus (CSFV). Computational modeling using AlphaFold 3 identified domain B of the E2 protein as the primary binding site for mAb 3E6. Building upon these findings, we established a cELISA employing mAb 3E6 and recombinant E2 protein. Receiver-operating characteristic (ROC) analysis revealed outstanding diagnostic performance, achieving a sensitivity of 99.26% and specificity of 98.99%. Further tests confirmed the cELISA's specificity for detecting BVDV-specific antibodies, with no cross-reactivity with antisera from animals infected or immunized against BCoV, BHV-1, BRV, AKAV, LSDV, BLV, and CSFV. Consistency was observed between results from the BVDV E2 cELISA and traditional virus neutralization test (VNT), demonstrating high sensitivity for monitoring antibody dynamics. In performance evaluations, the established cELISA exhibited high concordance with VNT in assessing 160 vaccinated sera and 190 clinical samples.DiscussionThe BVDV E2 cELISA, utilizing mAb 3E6 to target domain B of the BVDV E2 protein, represents a reliable and effective serological diagnostic tool for the detection of antibodies against both BVDV-1 and BVDV-2. This methodology holds significant promise for applications in clinical diagnosis and the evaluation of vaccine efficacy.

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Preclinical Efficacy of Cap-Dependent and Independent mRNA Vaccines against Bovine Viral Diarrhea Virus-1

Huang,

Hu,

Niu

et al. 2024

Veterinary Sciences

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Bovine viral diarrhea virus (BVDV) is an RNA virus associated with severe economic losses in animal production. Effective vaccination and viral surveillance are urgent for the prevention and control of BVDV infection. However, the application of traditional modified live vaccines and inactivated vaccines is faced with tremendous challenges. In the present study, we describe the preclinical efficacy of two BVDV mRNA vaccines tested in mice and guinea pigs, followed by a field trial in goats, where they were compared to a commercial vaccine (formaldehyde inactivated). The two mRNAs were engineered to express the envelope protein E2 of BVDV-1, the most prevalent subtype across the world, through a 5′ cap-dependent or independent fashion. Better titers of neutralizing antibodies against BVDV-1 were achieved using the capped RNA in the sera of mice and guinea pigs, with maximum values reaching 9.4 and 13.7 (by −log2), respectively, on the 35th day post-vaccination. At the same time point, the antibody levels in goats were 9.1 and 10.2 for the capped and capless RNAs, respectively, and there were no significant differences compared to the commercial vaccine. The animals remained healthy throughout the experiment, as reflected by their normal leukogram profiles. Collectively, our findings demonstrate that mRNA vaccines have good safety and immunogenicity, and we laid a strong foundation for the further exploitation of efficient and safe BVDV vaccines.

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Development and evaluation of a monoclonal antibody-based blocking ELISA to detect antibodies against the E2 protein of bovine viral diarrhea virus-1

Cited by 3 publications

References 37 publications

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

Development and Application of a Rapid Test for Detection of Bovine Viral Diarrhea Virus-specific Antibodies

Development of a pan-genotypic monoclonal antibody-based competitive ELISA for the detection of antibodies against Bovine viral diarrhea virus

Preclinical Efficacy of Cap-Dependent and Independent mRNA Vaccines against Bovine Viral Diarrhea Virus-1

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