Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul, 03722, Republic of Korea.
Department of Veterinary Medicine Virology Laboratory, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, 08826, Republic of Korea.
Small. 2023 Jun;19(26):e2207117. doi: 10.1002/smll.202207117. Epub 2023 Mar 24.
African swine fever virus (ASFV) is a severe and persistent threat to the global swine industry. As there are no vaccines against ASFV, there is an immense need to develop easy-to-use, cost-effective, and rapid point-of-care (POC) diagnostic platforms to detect and prevent ASFV outbreaks. Here, a novel POC diagnostic system based on affinity column chromatography for the optical detection of ASFV is presented. This system employs an on-particle hairpin chain reaction to sensitize magnetic nanoclusters with long DNA strands in a target-selective manner, which is subsequently fed into a column chromatography device to produce quantitatively readable and colorimetric signals. The detection approach does not require expensive analytical apparatus or immobile instrumentation. The system can detect five genes constituting the ASFV whole genome with a detection limit of ≈19.8 pm in swine serum within 30 min at laboratory room temperature. With an additional pre-amplification step using polymerase chain reaction (PCR), the assay is successfully applied to detect the presence of ASFV in 30 suspected swine samples with 100% sensitivity and specificity, similar to quantitative PCR. Thus, this simple, inexpensive, portable, robust, and customizable platform for the early detection of ASFV can facilitate the timely surveillance and implementation of control measures.
非洲猪瘟病毒 (ASFV) 是全球养猪业的严重且持久的威胁。由于目前尚无针对 ASFV 的疫苗，因此非常需要开发易于使用、具有成本效益且快速的即时检测 (POC) 诊断平台，以检测和预防 ASFV 爆发。在这里，提出了一种基于亲和柱色谱法的新型 POC 诊断系统，用于光学检测 ASFV。该系统采用基于颗粒的发夹链反应以目标选择性方式敏化磁性纳米簇与长 DNA 链，随后将其送入柱色谱设备以产生可定量读取和比色的信号。该检测方法不需要昂贵的分析仪器或固定的仪器。该系统可以在实验室室温下 30 分钟内检测到 ≈19.8 pm 的猪血清中构成 ASFV 全基因组的五个基因。通过聚合酶链反应 (PCR) 的额外预扩增步骤，该检测方法成功地应用于检测 30 个疑似猪样本中 ASFV 的存在，其灵敏度和特异性均为 100%，与定量 PCR 相似。因此，这种简单、廉价、便携、强大且可定制的 ASFV 早期检测平台可以促进及时监测和实施控制措施。
