Figure captions
Figure 1 :Typical clinical signs and pathological changes were
observed in PRV-infected goats, such as skin itch and fidgety
(A&B ), diarrhea (C ). Also, severe hyperemia and
hemorrhage were shown on the cerebrum from the infected goat
(D ).
Figure 2 : Isolation and identification of YNG strain from goat.
(A ) Obvious the cytopathic effects (CPE) of PK15 cells induced
by the infection of the third passages of YNG strain, the CPE of cells
in the left were characterized by being rounded and floated. The right
cells were set as negative group, Scale bar = 200 um. (B ) The
existence of infectious PRV virions in the cell cultures of the first,
second, third, fifth, and seventh passages were confirmed via PCR, which
could yielded the expected DNA bands (~450 bp) of PRV gE
in 1% agarose gel electrophoresis. DNA+ and DNA– were regarded as
positive and negative controls during DNA extraction, respectively. And
PCR- was the negative control during PCR amplification. (C )
Indirect immunofluorescent assay (IFA) for the detection of PRV in Vero
cells using anti-PRV gE and gB primary antibodies. Scale bar = 100 um
Figure 3 : The proliferation characteristics of YNG strains in
vitro. (A ) One-step growth curves of YNG and YNP strains on
PK15 and Vero cells at MOI of 0.01, the total PRV copies from the
supernatant and cells in triplicates were measured via real-time PCR.
(B ) The viral titers of YNG and YNP at 36 hpi were determined
in PK15 cells by TCID50.
Figure 4: Analysis of sequence variation in YNG strain.
(A ) A 7-aa insertion
(64AASTPAA70) in the gC gene of PRV
strains in genotype II compared with that in genotype I. (B ) A
2-aa deletion (278SP279) in the gD
gene of PRV variants in genotype II and Becker strain compared with that
of classical PRV strains in genotype II. (C ) A 3-nt deletion
(1469ACG1471) encoding D at the
position of 490 in the gE gene of YNG strain and another variant
PRV strain prevalent in China (GenBank no. MK806387 (JX/CH/2016)
compared with that of the other strains. (D&E ) Phylogenetic
trees based on gC and gE gene sequences of YNG strain and
other reference strains generated by the neighbor-joining method in MEGA
7.0 software. The red and black prismatic represented YNG strain
obtained in this study and PRV strain isolated from human, respectively.
Figure 5 : The pathogenicity of YNG strain in mouse model.
(A ) Survival curve (n=6/group) in Kunming mice after
intramuscular injection with different PRV strains
(104 TCID50/mouse). (B )
Pathological changes of cerebrum from mouse in different groups.
(C ) The viral copies in different organs from mice infected
with different PRV strains were determined according to the standard
curve constructed by the threshold cycle (CQ) values of the serial
10-fold dilution of standard plasmid with known copies via real-time PCR
method. Values are means±standard deviation (SD) = 3, significant
differences were determined when P>0.05 (ns),
P<0.05(*), P<0.01(**), and P<0.001(***).