INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Mengxiang Wang,Jinxing Song,Yanze Li,Junru Sun,Lei Zhou,Zhuoya Sun,Angke Zhang,Guoqing Zhuang,Pingan Xia,Gaiping Zhang,Yanan Wu
African swine fever has caused huge losses to the global pig industry. In the absence of effective vaccines, reliable detection methods are crucial. The p30 protein of ASFV is often used as a target for detection due to its high antigenicity in the early stage of virus replication. In this study, we modified the p30 protein using magnetic nanoparticles to increase its specific surface area and suspension in solution. Based on this modified p30 protein, we optimized the experimental steps to determine the best reaction conditions and then established a double probe competitive ELISA method. The established detection method can produce results within 45 min, with repeatability between and within batches <15 %, and it does not react with other pathogens, which demonstrates high specificity. In conclusion, we modified the ASFV p30 protein with magnetic nanoparticles and established a rapid, ultra-sensitive, and convenient detection method for ASF based on the modified p30. This also provides a new reference for the rapid detection of other veterinary diseases.
研究领域:
兽医学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ARCHIVES OF VIROLOGY
·
2021
·Vol. 166
·期 3
·871-879
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IF 2.574
Zhang Xinyu,Liu Xiaoyu,Wu Xiaodong,Ren Weijie,Zou Yanli,Xia Xiaoli,Sun Huaichang
African swine fever (ASF), caused by African swine fever virus (ASFV), was first reported in Kenya in 1921, but an effective vaccine or antiviral drug is still not available for ASFV control. Rapid and effective diagnostics are key steps in managing ASF. We generated two monoclonal antibodies (MAbs) against the ASFV phosphoprotein P30 and designated these as 3H7A7 and 6H9A10. Epitope mapping revealed that MAb 3H7A7 and 6H9A10 recognized aa 144-154 and aa 12-18 of P30, respectively. A signal-amplified sandwich colloidal gold test strip for rapid detection of ASFV was developed based using these MAbs. Sensitivity and specificity analysis showed that the detection limit of the strip was 2.16 ng of P30. The strip only reacted with ASFV and did not react with other common porcine viruses. In detection tests using 153 clinical field samples including sera, plasma, anticoagulant-treated blood, and tissue, the strip had 95.42% concordance with real-time PCR. The new MAbs specific for P30 and the rapid colloidal gold test strip helped to reveal novel B cell epitopes in P30 and provide an efficient diagnostic test for on-site clinical detection of ASF.
研究领域:
兽医学
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分子生物学
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免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
AMB Express
·
2025
·Vol. 15
·期 1
·1-12
·
IF 3.7
Hu Yanli,Xia Yingjun,Mo Huiwen,Zhang Li,Cao Wenjian,Fang Rui,Zhao Junlong
African swine fever (ASF) is a contagious viral disease in pigs, caused by the African swine fever virus (ASFV). Currently, there are no effective vaccines and drugs for ASFV, making diagnostic methods crucial for the prevention and control of ASF. In this study, the ASFV p30 protein was successfully expressed using a prokaryotic expression system and used as an immunogen to prepare monoclonal antibody (mAb) 2A5. The antigenic epitope recognized by the mAb 2A5 was identified as 58 VKYDIVKSARIYAGQGY 74 . Then, a blue latex microsphere immunochromatographic strip method for detecting ASFV antigens was established. Wherein a rabbit polyclonal antibody (pAb) against p30 stored in our laboratory was coupled with blue latex microspheres to prepare an immune probe, with mAb 2A5 as the test line and goat anti-rabbit IgG as the control line. The immunochromatographic strip exhibited excellent sensitivity, with a minimum detection limit of 10 ng/mL for the p30 protein. It demonstrated good specificity, showing no cross-reactivity with PRRSV, PCV2, PPV, and PEDV. Moreover, it maintained good stability and could be stored for at least 100 days at 37 °C. The concordance rate between the immunochromatographic strip and qPCR test kit was 92.2%. The immunochromatographic strip method can meet the requirements of clinical testing and provide technical support for ASFV diagnosis.
研究领域:
兽医学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Yuhao Huang,Huihui Li,Jian Wen,Shaowen Li,Qigai He,Xianrong Meng
Currently, antigen and antibody early detection is an important part in the control of African swine fever. However, traditional detection technologies based on antigen-antibody specific binding face problems such as low sensitivity, difficulty in preparing antibodies, and significant differences between batches of antibodies, which require the search for a new type of diagnostic molecule and detection technology. This study employed magnetic beads-SELEX to identify aptamers that specifically bind to the African swine fever virus (ASFV) p30 protein. The aptamer (p30-1) has high affinity and specificity, with no cross-reactivity to His-tagged or GST tagged proteins. In addition, the thiol-modified aptamer was used to immobilize ASFV p30 protein onto an electrochemical immunosensor constructed with thiol-modified multi-walled carbon nanotubes (S-MWCNTs) and gold nanoparticles (AuNPs). The detection of ASFV p30 monoclonal antibody was carried out using differential pulse voltammetry (DPV), and a standard curve was established. The linear detection range was 0.1 μg/mL-4 μg/mL, with a limit of detection (LOD) of 1.38 μg/mL. This method shows good specificity and reproducibility, enabling effective detection of clinical serum samples and infected animal serum samples. It provides new technical support for the rapid detection of ASFV antibodies.
研究领域:
兽医学
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分子生物学
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生物传感技术
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Lele Gong,Yuanyuan Zhang,Lele Wang,Xuyang Zhao,Lucai Wang,Xiangqi Qiu,Xilong Yang,Wenhui Zhu,Lijie Lv,Yunzhe Kang,Yanan Wu,Angke Zhang,Yongkun Du,Xuannian Wang,Gaiping Zhang,Aijun Sun,Guoqing Zhuang
The African swine fever virus (ASFV) continues to pose significant economic and pandemic risks. Consequently, discovering new, efficient vaccines is crucial. Messenger RNA (mRNA) vaccines have emerged as promising candidates, providing minimal risk of insertional mutagenesis, high safety profiles, effectiveness, rapid scalability in production, and cost-effectiveness. In this study, we have developed an ASF p30 mRNA vaccine candidate (mRNA/Man-LNP) employing mannose-modified lipid nanoparticles (LNPs). The mRNA/Man-LNP exhibited effective antigen presentation and facilitated dendritic cells (DCs) maturation. Notably, it elicited strong IgG titers and activated CD4 + and CD8 + T-cells in immunized mice, all while adhering to stringent biosafety standards. This investigation demonstrates that mRNA/Man-LNP can trigger both humoral and cellular immune responses, suggesting its potential as a potent and promising vaccine candidate for controlling African swine fever (ASF).
研究领域:
兽医学
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免疫学
·
分子生物学
·
病毒学
✓ 已验证
DOI
PubMed
高影响因子
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