Date Received: 03-04-2026 / Date Accepted: 09-06-2026 / Date Published: 26-10-2026
African swine fever (ASF) is a highly contagious disease with mortality rates of up to 100%. ASF virus (ASFV) can persist in livestock wastewater; however, its concentration is often low and affected by PCR inhibitors, posing challenges for detection. This study aimed to refine a polyethylene glycol (PEG)-based method for ASFV concentration before DNA extraction to improve detection efficiency using real-time PCR. The results identified optimal processing conditions, including a sample volume of 15ml, pre-centrifugation at 2,000g for 10 minutes, and the use of the supernatant for virus concentration. The optimal PEG 6000 concentration was 15% (w/v), with virus pelleting performed at 10,000g for 30 minutes. Two DNA extraction methods yielded comparable performance. Application of the PEG-based concentration protocol significantly improved ASFV detection compared to untreated samples. In field samples, the number of positive detections increased from 1/56 (untreated) to 8/56 (PEG-treated). These findings suggest that PEG-based virus concentration has strong potential for environmental surveillance and early detection of ASFV in pig production systems.
