Rapid Visible Detection of African Swine Fever Virus Using Hybridization Chain Reaction‐Sensitized Magnetic Nanoclusters and Affinity Chromatography

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. A rapid and accurate diagnostic system for African swine fever is developed using enzyme‐ and thermal recycling‐free on‐particle hybridization chain reaction and affinity column chromatography‐based signal detection. This strategy allows fast and easy target isolation, enzyme‐free DNA enrichment, and naked‐eye detection, which are highly beneficial for cost‐effective and field‐ready point‐of‐care testing of infectious diseases.