African Swine Fever

CAL Oura, L Edwards and CA Batten. 2013. Virological diagnosis of African swine fever—Comparative study of available tests. (Ask Eric for this paper.)

G Woźniakowski, M Frączyk, A Kowalczyk, M Pomorska-Mól, K Niemczuk and Z Pejsak. 2017. [Polymerase cross-linking spiral reaction (PCLSR) for detection of African swine fever virus (ASFV) in pigs and wild boars.]

• Polymerase cross-linking spiral reaction (PCLSR) is a method of amplifying targeted regions of DNA and can be used as a low cost way to check for the presence of viral DNA. Wozniakowski, et al. amplified a region of the p72 gene from African Swine Fever Virus. The reaction is optimized when performed at 65℃ steadily for 45 minutes. The procedure requires GspSSD polymerase for amplification, SYBR Green® I to fluorescently label the double stranded DNA product, and 2 sets of modified primers (sequences specific to ASFV included in methods). The primers include 3 major regions: 1 set covers the target region, 1 set flanks the target region, and 1 set protects the DNA in early steps and initiates cross-linked spiraling at the end. The third set originated from an unrelated Latrodectus hesperus gene designed to bind to itself but not the region of ASFV DNA. The estimated cost of each reaction is 2€ ($2.36).