Developing The Recombinase Polymerase Amplification (Rpa) Reaction To Detect The P30 Gene Of African Swine Fever (Asfv).

Abstract

African Swine Fever is a dangerous infectious disease that spreads rapidly and occurs in all species of pigs (including domestic and wild pigs). The disease occurs in all ages and all types of pigs, causing serious damage with a high fatality rate of up to 100%. In recent years, millions of pigs have been culled following disease outbreaks, impacting several sectors of the national economy and costing the government and farmers a substantial amount of money. The ASF virus has a high level of environmental resistance. Furthermore, pigs who recover from the illness could harbor the virus for an extended period or could be its permanent host, making it challenging to eradicate the virus in the event of an ASF outbreak. In addition, research for a vaccine or medical therapy to prevent this illness is currently lacking. Since the current biosecurity precautions are primarily preventive, a prompt laboratory diagnosis is required, and the pig herd must be eliminated if it is infected. PCR is now the most common technique for ASFV detection because of its sensitivity and effectiveness. The PCR method's sole disadvantage is that it necessitates the use of a thermocycler, which may be costly and time-consuming given that the reaction usually takes more than an hour to complete. To get over these drawbacks, the RPA assay was created to easily and swiftly detect ASFV. In this project, to identify ASFV, the p30 gene was amplified using specific primers, and the sample of DNA isolated from pig whole blood was subjected to RPA testing under one hour at 39°C. This served as the foundation for developing the colorimetric RPA test and offered a quick and effective way to detect ASFV.