2.2 Identification of PRV in collected specimens
The homogenates of each tissue specimen from goats or fecal samples from pigs living nearby were mixed with sterile phosphate-buffered saline (PBS) and undergone repeated freezing and thawing cycles, then centrifuged at 12000×g for 10 min. The viral nucleic acids in the supernatants were extracted using a commercial DNA/RNA extraction kit (Takara, Dalian, China), and stored at -80 ℃.
Prepared DNA samples were used for detecting the presence of PRV by PCR method with a pair of primers listed in Tab. 1 , PCR reaction (25 µL) mixture included 12.5 µL of 2×Taq Plus PCR Master mix (Takara, Dalian, China), 1 µL of each primer (10 pmol), 4 µL of DNA template, and 6.5 µL of DPEC treated water, the cycling parameters were performed as described previously (Tan et al., 2020). Prepared RNA samples were exploited to detect Rabies virus (RV) and Caprine arthritis encephalitis virus (GAEV), respectively, as previously established methods (Li et al., 2013). PCR products were visualized in 1% agarose gel electrophoresis.