Development and optimization of sampling techniques for environmental samples from African swine fever virus-contaminated surfaces with no organic contaminants.

African swine fever (ASF) is a highly contagious diseases in domestic pigs and wild boars with up to 100% mortality. ASF virus (ASFV) is a causative agent responsible for ASF and highly resistant in environments, which creates a significant challenge for the control and eradication of the virus. Despite the geographical expansion of ASFV and international movement of products to sustain the swine production system, there is limited knowledge on the use of environmental samples to perform surveillance to prevent the introduction of ASFV into ASFV-free areas and for control of transmission in affected areas. Therefore, this study aimed to develop and optimize sampling techniques for environmental samples for ASFV detection. The stainless steel surfaces were contaminated with ASFV-infected blood, swabbed using different devices, and then processed through different techniques. The environmental samples were processed and tested using qPCR analysis. The results showed that the use of pre-moistened gauze surgical sponges, sweeping pads, and sponge sticks resulted in increased sensitivity, when compared to either dry sampling devices or Dacron swab. In particular, the combination of the sponge stick and the commercial nucleic acid preservative supported the best detection of ASFV DNA on the clean stainless steel surfaces evaluated. Pre-incubation for the short period of time and centrifugation at low speed were sufficient to provide satisfactory diagnostic sensitivity of ASFV detection using qPCR for environmental samples. Our findings contribute to the development of techniques for environmental samples for ASFV surveillance to prevent the introduction and dissemination of ASFV.