Development and application of a high‐sensitivity immunochromatographic test strip for detecting classical swine fever virus antibodies

2021 ◽  
Author(s):  
Yilin Bai ◽  
Rui Jia ◽  
Qiang Wei ◽  
Li Wang ◽  
Yaning Sun ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
High Sensitivity ◽  
Test Strip ◽  
Fever Virus ◽  
Immunochromatographic Test

scholarly journals Development and Application of a High Sensitivity Immunochromatographic Test Strip for Detecting Classical Swine Fever Virus Antibody

2021 ◽  
Author(s):  
Yilin Bai ◽  
Rui Jia ◽  
Qiang Wei ◽  
Li Wang ◽  
Yaning Sun ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Clinical Sample ◽  
Expression System ◽  
Classical Swine Fever ◽  
Test Strip ◽  
Fever Virus ◽  
Control Measures ◽  
Antibody Detection ◽  
Immunochromatographic Test ◽  
E2 Protein

Abstract Background: Classical swine fever (CSF) is caused by classical swine fever virus (CSFV) and has lead huge losses to the pig industry worldwide. Although vaccination and other control measures have been carried out, it is essential to establish a rapid valid method for CSF vaccination monitoring and clinical diagnosis. CSFV E2 protein has been well known as a major antigen for antibody detection. It is significant to improve affinity between E2 protein and CSFV antibody to result in a better performance of detection method. Results: In this study, a recombinant E2 extracellular protein (aa 1-331), which was with the native homodimer conformation and had a high affinity with anti-CSFV-E2 monoclonal antibody WH303, was expressed using Bac-to-Bac baculovirus expression system. A novel immunochromatographic test strip based on the recombinant CSFV E2 protein was developed for CSFV antibody detection. The sensitivity of this strip for detecting CSFV standard positive serum was 1:102400, 4 times higher than that of the former developed CnC2 test strip. No cross reaction with other swine virus antibodies was observed. The detection of clinical swine serum samples (n=138) demonstrated that the agreement of the new E2 test strip with three commercial ELISA kits was 88.40% (122/138), 86.23% (119/138), and 96.38% (133/138), respectively. Conclusion: Our data indicate that a novel E2 test strip with higher sensitivity has been developed and can be applied for clinical sample detections, providing a new powerful and simple approach for future monitoring CSFV antibodies.

scholarly journals Development and application of a high sensitivity immunochromatographic test strip for detecting classical swine fever virus antibody

2021 ◽  
Author(s):  
Yilin Bai ◽  
Rui Jia ◽  
Qiang Wei ◽  
Li Wang ◽  
Yaning Sun ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Clinical Sample ◽  
Expression System ◽  
Classical Swine Fever ◽  
Test Strip ◽  
Fever Virus ◽  
Control Measures ◽  
Antibody Detection ◽  
Immunochromatographic Test ◽  
E2 Protein

Classical swine fever (CSF) is caused by classical swine fever virus (CSFV) and has led to huge ecnomic losses for the pig industry worldwide. Although vaccination and other control measures have been carried out, it is essential to establish a rapid and valid method for CSF vaccination monitoring and clinical diagnosis. CSFV E2 protein has been well-known as a major antigen for antibody detection. It is significant to improve affinity between E2 protein and CSFV antibody for a better performance of detection method. In this study, a recombinant E2 extracellular protein (aa 1-331), which has a native homodimer conformation and has a high affinity with anti-CSFV-E2 monoclonal antibody WH303, was expressed using Bac-to-Bac baculovirus expression system. A novel immunochromatographic test strip based on the recombinant CSFV E2 protein was developed for CSFV antibody detection. The sensitivity of this strip for detecting CSFV standard positive serum was 1:102400, 4 times higher than that of the previously developed CnC2 test strip. No cross reaction with antibodies of other swine viruses was observed. The detection of clinical swine serum samples (n=138) demonstrated that the agreements of this E2 test strip with three commercial ELISA kits were 88.40% (122/138), 86.23% (119/138), and 96.38% (133/138), respectively. Our data indicated that a novel E2 test strip with higher sensitivity has been developed and can be applied for clinical sample detections, providing a new powerful and simple approach for CSFV antibody monitoring.

scholarly journals Development and Application of a RT-NestPCR Assay for Differential Detection of C-Strain and Wild-Type Viruses of Classical Swine Fever Virus

2013 ◽  
Vol 2 (3) ◽  
pp. 27
Author(s):  
Haiguang Wang ◽  
Yibao Ning ◽  
Cong Ying ◽  
Can Liu ◽  
Ruirui Wei ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Gel Electrophoresis ◽  
Classical Swine Fever ◽  
High Sensitivity ◽  
Fever Virus ◽  
Test Method ◽  
Wild Type Virus ◽  
Diagnostic Tools ◽  
Differential Detection ◽  
Wild Type

<p>Due to the urgent need of differentiation of infected from vaccinated animals in control and eradication of classical swine fever (CSF) and the shortcomings of current differential diagnostic tools, this study is aiming to establish a RT-nestPCR assay for differential detection of wild-type viruses and lapinized Chinese vaccine strain (C-strain) of classical swine fever virus (CSFV) of high sensitivity. Two pairs of CSFV-specific primers were designed in the conservative regions of NS5B (a non-structural protein encoded by the CSFV genome, which performs the RNA dependent RNA polymerase activity) and 3? un-translated regions (3?-UTR) to encompass the T-rich insertion uniquely existing in the 3?-UTR of C-strain genome. Thus the amplification fragment of C-strain is longer than that of the wild-type viruses for it contains the T-rich insertion region. Two pairs of primers were used in combination and the wild-type viruses and C-strain of CSFV could be detected and accurately distinguished with a high sensitivity through super fine resolution (SFR) argarose gel electrophoresis that displays the different lengths of the amplicons. The detection limit of the C-strain and Shimen strain were respectively 4.5×10<sup>-2 </sup>pg and 3.2×10<sup>-2 </sup>pg of viral RNA. The results of the specificity test showed that this method can detect different strains of CSFV without amplifying other non-CSFV pathogens. The results of the detection of 400 clinical samples indicated that 16 samples were CSFV positive in total; in which 4 samples were C-Strain positive and 12 were wild-type CSFV positive. The total CSFV positive rate was 4%. The detection results of the 14 batches of C-Strain vaccines showed that all samples displayed bands of C-Strain amplicons in the SFR argarose gel electrophoresis and all vaccines were free of wild-type virus contamination. In conclusion, the RT-nestPCR assay established in the present study could supply a sensitive and specific test method for distinguishing wild-type CSFV infected animals from those vaccinated with C-strain vaccines in the field.</p>

Purification of Classical Swine Fever Virus E2 Subunit Vaccines basing on High Affinity Peptide Ligand.

2020 ◽  
Vol 27 ◽  
Author(s):  
Fangyu Wang ◽  
Qiuying Yu ◽  
Man Hu ◽  
Guangxu Xing ◽  
Dong Zhao ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Rapid Development ◽  
Classical Swine Fever ◽  
High Sensitivity ◽  
Fever Virus ◽  
Affinity Constant ◽  
Peptide Ligand ◽  
Binding Interaction ◽  
Sensitivity And Selectivity

Background: The purification of expressed proteins is the most critical part of subunit-vaccine production. Protein-purification methods such as affinity chromatography and ion exchange still have the shortcomings of being time consuming and complicated. With the rapid development of computational molecular-simulation technology, structure-based peptide-ligand design has become feasible. Objection: We aimed to apply molecular docking for a peptide ligand designed for classical swine fever virus (CSFV) E2 purification. Methods: Computational-derived peptides were synthesized, and the in vitro binding interaction with E2 was investigated. The effects of purification on E2 were also evaluated. Results: The best peptide recognizing E2 was P6, which had a sequence of KKFYWRYWEH. Based on kinetic surface plasmon resonance (SPR) analysis, the apparent affinity constant of P6 was found to be 148 nM. Importantly, P6 showed suitable binding affinity and specificity for E2 purification from transgenic rice seeds. Evaluation of immune antibodies in mice showed that the antibody-blocking rate on day 42 after inoculation reached 86.18% and 90.68%. Conclusion: The computational-designed peptide in this study has high sensitivity and selectivity and is thus useful for the purification of CSFV E2. The novel method of design provided a broad platform and powerful tool for protein-peptide screening, as well as new insights into CSFV vaccine design.

ARFGAP1 binds to classical swine fever virus NS5A protein and enhances CSFV replication in PK-15 cells

2021 ◽  
Vol 255 ◽  
pp. 109034
Author(s):  
Liang Zhang ◽  
Mingxing Jin ◽  
Mengzhao Song ◽  
Shanchuan Liu ◽  
Tao Wang ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Ns5a Protein

Genome Variability of Classical Swine Fever Virus during the 2018–2020 Epidemic in Japan

2021 ◽  
pp. 109128
Author(s):  
Tatsuya Nishi ◽  
Katsuhiko Fukai ◽  
Tomoko Kato ◽  
Kotaro Sawai ◽  
Takehisa Yamamoto
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Genome Variability

scholarly journals A Novel E2 Glycoprotein Subunit Marker Vaccine Produced in Plant Is Able to Prevent Classical Swine Fever Virus Vertical Transmission after Double Vaccination

Vaccines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 418
Author(s):  
Youngmin Park ◽  
Yeonsu Oh ◽  
Miaomiao Wang ◽  
Llilianne Ganges ◽  
José Alejandro Bohórquez ◽  
...  
Keyword(s):  
Vertical Transmission ◽  
Classical Swine Fever Virus ◽  
Neutralizing Antibodies ◽  
Subunit Vaccine ◽  
Vaccine Dose ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Tissue Samples ◽  
Marker Vaccine ◽  
E2 Glycoprotein

The efficacy of a novel subunit vaccine candidate, based in the CSFV E2 glycoprotein produced in plants to prevent classical swine fever virus (CSFV) vertical transmission, was evaluated. A Nicotiana benthamiana tissue culture system was used to obtain a stable production of the E2-glycoprotein fused to the porcine Fc region of IgG. Ten pregnant sows were divided into three groups: Groups 1 and 2 (four sows each) were vaccinated with either 100 μg/dose or 300 μg/dose of the subunit vaccine at 64 days of pregnancy. Group 3 (two sows) was injected with PBS. Groups 1 and 2 were boosted with the same vaccine dose. At 10 days post second vaccination, the sows in Groups 2 and 3 were challenged with a highly virulent CSFV strain. The vaccinated sows remained clinically healthy and seroconverted rapidly, showing efficient neutralizing antibodies. The fetuses from vaccinated sows did not show gross lesions, and all analyzed tissue samples tested negative for CSFV replication. However, fetuses of non-vaccinated sows had high CSFV replication in tested tissue samples. The results suggested that in vaccinated sows, the plant produced E2 marker vaccine induced the protective immunogenicity at challenge, leading to protection from vertical transmission to fetuses.

Characterization of antibody responses against a neutralizing epitope on the glycoprotein E2 of classical swine fever virus

2008 ◽  
Vol 153 (8) ◽  
pp. 1593-1598 ◽  
Author(s):  
Y. Qi ◽  
L. C. Liu ◽  
B. Q. Zhang ◽  
Z. Shen ◽  
J. Wang ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Antibody Responses ◽  
Neutralizing Epitope ◽  
Glycoprotein E2

scholarly journals Classical swine fever virus employs the PERK- and IRE1-dependent autophagy for viral replication in cultured cells.

Virulence ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 130-149
Author(s):  
Erpeng Zhu ◽  
Huawei Wu ◽  
Wenxian Chen ◽  
Yuwei Qin ◽  
Jiameng Liu ◽  
...  
Keyword(s):  
Viral Replication ◽  
Classical Swine Fever Virus ◽  
Cultured Cells ◽  
Classical Swine Fever ◽  
Fever Virus

scholarly journals Research Article An improved Bac-to-Bac/BmNPV technology expressing envelope E2 glycoprotein of classical swine fever virus (CSFV) in the silkworm, Bombyx mori

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
X. Hu ◽  
Y. Chen ◽  
C. O. Singh ◽  
S. Zhang ◽  
Y.G. Miao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Research Article ◽  
E2 Glycoprotein
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