School of Life Sciences, Zhengzhou University, Zhengzhou, 450001, Henan, China; Longhu Laboratory, Zhengzhou, Henan, China.
School of Life Sciences, Zhengzhou University, Zhengzhou, 450001, Henan, China.
Int J Biol Macromol. 2024 Nov;279(Pt 4):134852. doi: 10.1016/j.ijbiomac.2024.134852. Epub 2024 Aug 17.
The African swine fever virus (ASFV), a highly contagious pathogen responsible for African swine fever (ASF), causes significant economic losses in the global pork industry. Due to its large and complex structure, ASFV remains refractory to commercial vaccine development, necessitating the creation of rapid, sensitive, and specific diagnostic tools for disease control. In this study, quantum dots were conjugated to ASFV p72 protein to establish a fluorescent immunochromatographic assay for detecting ASFV-specific antibodies. The assay test strips contained four adjacent pads arranged sequentially: a sample-application pad, a pad containing mobile antigen-probe conjugate, a nitrocellulose readout pad featuring a test line containing immobilised staphylococcal protein A and a control line containing immobilised monoclonal antibodies against the ASFV p72 protein, and an absorbent pad driving the directional flow of liquid via capillary action. The resulting fluorescence immunochromatographic assay demonstrated highly sensitive and specific ASFV antibody detection in under 15 min. Specificity testing showed no cross-reactivity with serum antibodies against other viruses and sensitivity surpassing that of commercial ASFV antibody colloidal gold immunochromatographic test strips. This novel approach offers rapid detection, excellent specificity, and high sensitivity, and supports the future development of fluorescent immunochromatographic test strips for ASFV antibody detection.
非洲猪瘟病毒(ASFV)是一种高度传染性病原体,可导致非洲猪瘟(ASF),给全球猪肉产业造成重大经济损失。由于其庞大而复杂的结构,ASF 仍然难以开发出商业疫苗,因此需要快速、敏感和特异性的诊断工具来进行疾病控制。在这项研究中,量子点与 ASFV p72 蛋白偶联,建立了用于检测 ASFV 特异性抗体的荧光免疫层析分析方法。检测试条包含四个相邻的垫,依次排列:样品垫、含有移动抗原-探针偶联物的垫、硝酸纤维素读出垫,其中包含固定化的葡萄球菌蛋白 A 的测试线和固定化针对 ASFV p72 蛋白的单克隆抗体的控制线,以及一个吸收垫,通过毛细作用驱动液体的定向流动。所得到的荧光免疫层析分析方法在 15 分钟内即可实现高灵敏度和特异性的 ASFV 抗体检测。特异性测试显示与针对其他病毒的血清抗体无交叉反应性,且灵敏度超过了商业 ASFV 抗体胶体金免疫层析检测试条。这种新方法具有快速检测、优异的特异性和高灵敏度,支持未来开发用于 ASFV 抗体检测的荧光免疫层析检测试条。