p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies
In this study, a novel rapid immunochromatographic (IC) test for African swine fever virus (ASFV) antibodies is presented. An immunochromatographic test (IC) is a detection technique that combines membrane chromatography with immunolabeling. This approach saves time for antibody preparation, resulti...
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2025-01-01
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| author | Yang Yang Yuhao Li Ziyang Wang Minglong Tong Pengcheng Zhu Juanxian Deng Zongjie Li Ke Liu Beibei Li Donghua Shao Zhongren Zhou Yafeng Qiu Zhiyong Ma Jianchao Wei |
| author_facet | Yang Yang Yuhao Li Ziyang Wang Minglong Tong Pengcheng Zhu Juanxian Deng Zongjie Li Ke Liu Beibei Li Donghua Shao Zhongren Zhou Yafeng Qiu Zhiyong Ma Jianchao Wei |
| author_sort | Yang Yang |
| collection | DOAJ |
| description | In this study, a novel rapid immunochromatographic (IC) test for African swine fever virus (ASFV) antibodies is presented. An immunochromatographic test (IC) is a detection technique that combines membrane chromatography with immunolabeling. This approach saves time for antibody preparation, resulting in a shorter production cycle. p54 is an important structural protein of African swine fever, and an ideal protein for serotype diagnosis. Gold nanoparticles are attached to the ASFV p54-Fc fusion protein, and the ASFV-specific antigen p54 and Staphylococcus aureus protein A (SPA) are labeled on a nitrocellulose membrane, at positions T and C, respectively. We developed a SPA double sandwich IC test strip, and assessed its feasibility using ASFV p54 and p54-Fc fusion proteins as antigens. ASFV p54 and p54-Fc fusion proteins were expressed and purified. A sandwich cross-flow detection method for p54, which is the primary structural protein of ASFV, was established, using colloidal gold conjugation. Our method can detect ASFV antibodies in field serum samples in about 15 min using a portable colloidal gold detector, demonstrating high specificity and sensitivity (1:320), and the coincidence rate was 98% using a commercial ELISA kit. The dilution of the serum sample can be determined by substituting the absorbance (T-line) interpreted by portable devices into the calibration curve function formula of an African swine fever virus standard serum. In summary, our method is rapid, cost-effective, precise, and highly selective. Additionally, it introduces a new approach for constructing IC test strips using SPA protein without antibody preparation, making it a reliable on-site antibody test for ASFV. |
| format | Article |
| id | doaj-art-e42b0a86068a4fe580b4bf51dd162e38 |
| institution | Kabale University |
| issn | 2079-6374 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj-art-e42b0a86068a4fe580b4bf51dd162e382025-01-24T13:25:28ZengMDPI AGBiosensors2079-63742025-01-011512510.3390/bios15010025p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV AntibodiesYang Yang0Yuhao Li1Ziyang Wang2Minglong Tong3Pengcheng Zhu4Juanxian Deng5Zongjie Li6Ke Liu7Beibei Li8Donghua Shao9Zhongren Zhou10Yafeng Qiu11Zhiyong Ma12Jianchao Wei13Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaYixing Customs, Yixing 214200, ChinaYixing Customs, Yixing 214200, ChinaNanjing Customs, Nanjing 210001, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Quicking Biotech Co., Shanghai 201314, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, ChinaIn this study, a novel rapid immunochromatographic (IC) test for African swine fever virus (ASFV) antibodies is presented. An immunochromatographic test (IC) is a detection technique that combines membrane chromatography with immunolabeling. This approach saves time for antibody preparation, resulting in a shorter production cycle. p54 is an important structural protein of African swine fever, and an ideal protein for serotype diagnosis. Gold nanoparticles are attached to the ASFV p54-Fc fusion protein, and the ASFV-specific antigen p54 and Staphylococcus aureus protein A (SPA) are labeled on a nitrocellulose membrane, at positions T and C, respectively. We developed a SPA double sandwich IC test strip, and assessed its feasibility using ASFV p54 and p54-Fc fusion proteins as antigens. ASFV p54 and p54-Fc fusion proteins were expressed and purified. A sandwich cross-flow detection method for p54, which is the primary structural protein of ASFV, was established, using colloidal gold conjugation. Our method can detect ASFV antibodies in field serum samples in about 15 min using a portable colloidal gold detector, demonstrating high specificity and sensitivity (1:320), and the coincidence rate was 98% using a commercial ELISA kit. The dilution of the serum sample can be determined by substituting the absorbance (T-line) interpreted by portable devices into the calibration curve function formula of an African swine fever virus standard serum. In summary, our method is rapid, cost-effective, precise, and highly selective. Additionally, it introduces a new approach for constructing IC test strips using SPA protein without antibody preparation, making it a reliable on-site antibody test for ASFV.https://www.mdpi.com/2079-6374/15/1/25immunochromatographic (IC) testSPAFc fusion proteinASFVfield rapid detection |
| spellingShingle | Yang Yang Yuhao Li Ziyang Wang Minglong Tong Pengcheng Zhu Juanxian Deng Zongjie Li Ke Liu Beibei Li Donghua Shao Zhongren Zhou Yafeng Qiu Zhiyong Ma Jianchao Wei p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies Biosensors immunochromatographic (IC) test SPA Fc fusion protein ASFV field rapid detection |
| title | p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies |
| title_full | p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies |
| title_fullStr | p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies |
| title_full_unstemmed | p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies |
| title_short | p54-Fc-Labeled Gold Nanoparticle-Based Lateral Flow Strip-Assisted Portable Devices for Rapid and Quantitative Point-of-Care Detection of ASFV Antibodies |
| title_sort | p54 fc labeled gold nanoparticle based lateral flow strip assisted portable devices for rapid and quantitative point of care detection of asfv antibodies |
| topic | immunochromatographic (IC) test SPA Fc fusion protein ASFV field rapid detection |
| url | https://www.mdpi.com/2079-6374/15/1/25 |
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