Detection Of Borna Diease Virus P24 Fragment And Antigen In Brain Tissue Of Bats

Objective: To investigate the infection situation of Borna disease virus(BDV)in wild bats and to enrich the epidemiology of BDV in Guizhou Province,and to lay the foundation for BDV prevention and control.Methods: Among April and September in 2014 the wild bats were captured by bird nets at Shuanghe Cave in Zunyi, Guizhou, and the bat frontotemporal brain tissues were taken under strict asepetic way. The p24 gene fragment of BDV-RNA in brain tissue of bat was detected by nested reverse transcriptase polymerase chain reaction(n RT-PCR). The positive n RT-PCR products were analyzed by genetic sequence,homology and molecular phylogenetic tree. In addition the phosphoprotein expression of BDV in 82 wild bats brain tissue specimens were detected by Western Blot(WB).Results: It showed that the extended p24 gene fragments of BDV-RNA in brain tissue from 82 bats was 7 positived(22, 33, 36, 48, 55, 58, 68). Only 5positive products(22, 33, 36, 48, 58)could be successfully detected the gene sequences,the two samples(55, 68) were not detected the gene sequences because of the low concentration.Its positive percentage was 6.1%(5/82). All the amplified gene fragments were verified the BDV-p24 gene sequences(nt1528- nt 1750), but the BDV-p24 gene sequences fragments of 5 positive samples were not all the same. The nucleotide sequences homology of BDV-p24 gene fragment was 92%-99% when separately compared with standard Strain V in horse(Gene Bank number: AJ31152.1), H1766(Gene Bank number: AJ311523.1), He/80(Gene Bank number: AJ311522), especially the22 th positive samples, its homology was high up to 99%. The 36 th, 48 th, 58 th nucleotide mutations were mainly concentrated in the nt1734- nt1740 location on the BDV-p24 gene sequence and the nucleotide on this location of this three positive samples was GGAA(T)GAA. The point mutations of 33 th sample were occurred widely. The molecular phylogenetic tree found that the wild bats’ nucleotide sequences of positive specimens belonging to 2branches(4 BDV-p24 gene-positive samples were brought together to form independent branch in Zunyi, Guizhou Province and 1 BDV-p24gene-positive sample was gathered together to form Germany-China mixed lineage). The WB result showed that 9 of 82 specimens were phosphoprotein positive. Its positive percentage was 10.98%(9/82).Conclusion: This study suggest that the wild bats may be the carrier of BDV natural infection in Zunyi, Guizhou Province. It prompts that BDV can circulate communications between bats,bats and human.