Zhu Wenzhuang, Meng Kaiwen, Zhang Yueping, Bu Zhigao, Zhao Dongming, Meng Geng
College of Veterinary Medicine, China Agricultural University, Beijing, China.
State Key Laboratory of Veterinary Biotechnology, National High Containment Facilities for Animal Diseases Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
Front Chem. 2022 Jan 3;9:804981. doi: 10.3389/fchem.2021.804981. eCollection 2021.
African swine fever is a widespread and highly contagious disease in the porcine population, which is caused by African swine fever virus (ASFV). The PCR and ELISA detection methods are the main conventional diagnostic methods for ASFV antigen/antibody detection in the field. However, these methods have limitations of expensive equipment, trained technicians, and time-consuming results. Thus, a rapid, inexpensive, accurate and on-site detection method is urgently needed. Here we describe a double-antigen-sandwich lateral-flow assay based on gold nanoparticle-conjugated ASFV major capsid protein p72, which can detect ASFV antibody in serum samples with high sensitivity and specificity in 10 min and the results can be determined by naked eyes. A lateral flow assay was established by using yeast-expressed and acid-treated ASFV p72 conjugated with gold nanoparticles, which are synthesized by seeding method. A high coincidence (97.8%) of the assay was determined using clinical serum compared to a commercial ELISA kit. In addition, our lateral flow strip can detect as far as 1:10,000 diluted clinically positive serum for demonstration of high sensitivity. In summary, the assay developed here was shown to be rapid, inexpensive, accurate and highly selective. It represents a reliable method for on-site ASFV antibody detection and may help to control the ASFV pandemic.
非洲猪瘟是猪群中一种广泛传播且极具传染性的疾病，由非洲猪瘟病毒（ASFV）引起。聚合酶链式反应（PCR）和酶联免疫吸附测定（ELISA）检测方法是该领域检测ASFV抗原/抗体的主要传统诊断方法。然而，这些方法存在设备昂贵、需要训练有素的技术人员以及结果耗时等局限性。因此，迫切需要一种快速、廉价、准确且可现场检测的方法。在此，我们描述了一种基于金纳米颗粒偶联的ASFV主要衣壳蛋白p72的双抗原夹心侧流分析法，该方法可在10分钟内高灵敏度和特异性地检测血清样本中的ASFV抗体，且结果可用肉眼判定。通过使用酵母表达并经酸处理的与金纳米颗粒偶联的ASFV p72建立了一种侧流分析法，金纳米颗粒通过种子法合成。与商业ELISA试剂盒相比，使用临床血清测定该分析法的一致性较高（97.8%）。此外，我们的侧流试纸条可检测至临床阳性血清1:10,000稀释度，以证明其高灵敏度。总之，此处开发的分析法显示出快速、廉价、准确且高度选择性的特点。它代表了一种用于现场检测ASFV抗体的可靠方法，可能有助于控制ASFV的大流行。
