PDF(Pubmed)
Abstract：
African Swine Fever (ASF), caused by the African swine fever virus (ASFV), has inflicted significant economic losses on the pig industry in China. The key to mitigating its impact lies in accurate screening and strict biosecurity measures. In this regard, the development of colloidal gold immunochromatographic test strips (CGITS) has proven to be an effective method for detecting ASFV antibodies. These test strips are based on the ASFV p30 recombinant protein and corresponding monoclonal antibodies. The design of the test strip incorporates a high-concentration colloidal gold-labeled p30 recombinant protein as the detection sensor, utilizing Staphylococcal Protein A (SPA) as the test line (T line), and p30 monoclonal antibody as the control line (C line). The sensitivity and specificity of the test strip were evaluated after optimizing the labeling concentration, pH, and protein dosage. The research findings revealed that the optimal colloidal gold labeling concentration was 0.05 %, the optimal pH was 8.4, and the optimal protein dosage was 10 μg/mL. Under these conditions, the CGITS demonstrated a detection limit of 1:512 dilution of ASFV standard positive serum, without exhibiting cross-reactivity with antibodies against other viral pathogens. Furthermore, the test strips remained stable for up to 20 days when stored at 50 °C and 4 °C. Comparatively, the CGITS outperformed commercial ELISA kits, displaying a sensitivity of 90.9 % and a specificity of 96.2 %. Subsequently, 108 clinical sera were tested to assess its performance. The data showed that the coincidence rate between the CGITS and ELISA was 93.5 %. In conclusion, the rapid colloidal gold test strip provides an efficient and reliable screening tool for on-site clinical detection of ASF in China. Its accuracy, stability, and simplicity make it a valuable asset in combating the spread of ASF and limiting its impact on the pig industry.
摘要：
非洲猪瘟(ASF)由非洲猪瘟病毒（ASFV）引起，给中国养猪业造成了巨大的经济损失。减轻其影响的关键在于准确的筛查和严格的生物安全措施。在这方面,胶体金免疫层析试纸条（CGITS）的开发已被证明是检测ASFV抗体的有效方法。这些测试条基于ASFVp30重组蛋白和相应的单克隆抗体。试纸条的设计采用高浓度胶体金标记p30重组蛋白作为检测传感器,利用葡萄球菌蛋白A（SPA）作为测试线（T线），和p30单克隆抗体作为对照线(C线)。在优化标记浓度后,评价了试纸条的敏感性和特异性,pH值,和蛋白质剂量。研究结果表明,胶体金标记的最佳浓度为0.05%,最佳pH值为8.4,最佳蛋白用量为10μg/mL。在这些条件下,CGITS的检测限为1:512稀释的ASFV标准阳性血清，不表现出与针对其他病毒病原体的抗体的交叉反应性。此外，当在50°C和4°C下储存时,测试条保持稳定长达20天。相对而言,CGITS优于商业ELISA试剂盒，敏感性为90.9%，特异性为96.2%。随后，测试了108份临床血清以评估其性能。数据显示，CGITS与ELISA的符合率为93.5％。总之,快速胶体金试纸条为我国ASF的现场临床检测提供了高效可靠的筛选工具。其准确性,稳定性,和简单性使其成为打击ASF蔓延和限制其对养猪业影响的宝贵资产。
