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.