89%) and the nucleotide sequence ITF2357 nmr Identity was lowest between the YN08 isolate and the South Korean isolate (93.61%). Table 4 Percent Identity (below the diagonal) and Divergence (above the diagonal) matrix of 3′ UTR sequence of different Alphavirus isolates   1 2 3 4 5 6 7 8 9 10 1. ALPV_M1   0.0035 0.0046 0.0023 0.0011 0.0118 0.0626 0.0035 0.0035 0.0011 2. GETV_HB0234 99.65% GDC-0449 mouse   0.0082 0.0012 0.0023 0.0155 0.0640 0.0023 0.0023 0.0046 3. GETV_LEIV_16275_Mag 99.54% 99.18%   0.0070 0.0058 0.0142 0.0656 0.0082 0.0082 0.0058 4. GETV_LEIV_17741_MPR 99.77% 99.88% 99.30%   0.0012 0.0143 0.0625 0.0011 0.0012 0.0035 5. GETV_M1

99.89% 99.77% 99.42% 99.88%   0.0130 0.0641 0.0023 0.0023 0.0023 6. GETV_MM2021 98.82% 98.45% 98.58% 98.57% 98.70%   0.0781

0.0155 0.0155 0.0130 7. GETV_S_Korea 93.74% 93.60% 93.44% 93.75% 93.59% 92.19%   0.0639 0.0640 0.0626 8. GETV_YN08 99.65% 99.77% 99.18% 99.89% 99.77% 98.45% VX-689 93.61%   0.0023 0.0046 9. GETV_YN0540 99.65% 99.77% 99.18% 99.88% 99.77% 98.45% 93.60% 99.77%   0.0046 10.SAGV(DNA) 99.89% 99.54% 99.42% 99.65% 99.77% 98.70% 93.74 99.54% 99.54%   Phylogenetic analysis To better understand the genetic relationship of YN08 to other strains of Getah virus in the world (including Chinese isolates ALPV_M1, GETV_M1, HB0234, and YN0540), the previously published genetic sequences of GETV and other alphavirus capsid protein genes and 3’-UTR

sequences obtained from GenBank were used to construct phylogenetic trees. The phylogenetic analyses clearly showed that YN08 is more closely related to the Hebei HB0234 strain than the YN0540 strain, and more distantly related to the MM2021 Malaysia primitive strain (Figure 3). Figure 3 Phylogenetic nearly relationship betweenYN08 isolates of GETV and other alphaviruses based on the non-structural protein gene nsP3, capsid protein and 3′ UTR area sequences. The neighbor joining tree was constructed using the MEGA with bootstrapping. (A) Phylogenetic analysis of RT-PCR sequences of the non-structural protein gene nsP3 from YN08 isolates of GETV and other alphaviruses. (B) Phylogenetic tree constructed using the nucleotide sequences of the capsid gene of YN08 isolates of GETV and other alphaviruses. (C) Phylogenetic tree constructed using the nucleotide sequences of 3’-UTR area sequences of GETV isolates. Discussion Alphaviruses are mosquito-borne RNA viruses that cause devastating or debilitating diseases in both humans and livestock. SAGV and GETV are two members of the Alphavirus genus of the family Togaviridae. GETV is widely distributed in southeast Asia and northern Australia along the Pacific Ocean [20–24]. GETV has been isolated from various mosquito species of the genera Culex, Aedes, and Armigeres[18].