Polymerase cross-linking spiral reaction (PCLSR) for detection of African swine fever virus (ASFV) in pigs and wild boars.

摘要: The study reports the development of a polymerase cross-linking spiral reaction (PCLSR) for detection African swine fever virus (ASFV) DNA in blood collected from infected pigs and wild boars. method uses 3 specifically designed primers. Two outer-spiral primers comprising 3′ sequences complementary to ASFV p72 gene sequence 5′end exogenous black widow alpha-latrotoxin as well additional specific primer. is exclusively without cross-reactions with cDNA classical (CSFV), porcine reproductive respiratory syndrome (PRRSV) or epidemic diarrhea (PEDV). sensitivity this technique reached 7.2 × 102 copies per μl−1 plasmid containing gene. PCLSR was conducted at 65 °C creating cross-linked complex structures. results were visualized using SYBR Green I dye, gel electrophoresis while progress traced real-time PCR system that resulted registration fluorescent curves melting peaks 85.3 °C. developed examined tissue samples selected 17 ASF cases boars outbreaks pigs. Further tests have been also 55 23 22 cases. These showed might be further used preliminary cost-effective surveillance ASFV.