Comparison Of External Incubation Period Of Aedes Aegypti Infected With DENV?ZIKV And Infection Characteristics

Aedes aegypti is main vector of dengue and zika,it is important to verify whether there are differences in the external latency of Ae.aegypti infected with Dengue virus(DENV)and Zika virus(ZIKV),explore the characteristics of the differences and the possible mechanism of the differences,and find the related genes or pathways that affect the differences in the external latency,so as to provide scientific basis for the control of mosquito borne virus and the elucidation of the antiviral mechanism of mosquitoes.The research consisted of collection of Ae.aegypti and establishment of laboratory domestication,virus amplification and virus detection system,comparison of virus replication after Ae.aegypti cells infected with DENV and ZIKV,comparison of virus infection rate and replication after Ae.aegypti infected with DENV and ZIKV,and differential analysis and functional analysis of mi RNA and m RNA expression profiles between Ae.aegypti cells and Ae.aegypti tissues.The results are as follows,(1)Under the same experimental conditions,when Aedes aegypti Aag2 cells were infected with DENV-2 and ZIKV respectively,the initial amount of ZIKV entering Aag2 cells was higher than DENV-2.In the following days,ZIKV replication speed was faster than DENV-2,and the virus titer of ZIKV reached the peak 48 hours after infection,while DENV-2 slowly increased to the peak in 1-7days,until it was close to the virus droplet of ZIKV Degree level.(2)After Ae.aegypti was infected with DENV and zikv,the external incubation period of Ae.aegypti infected with ZIKV was shorter than that of Ae.aegypti infected with DENV-2,and the virus infection rate and virus copy number in tissues of Ae.aegypti infected with ZIKV were higher than those of Ae.aegypti infected with DENV.(3)In the sequencing results of Ae.aegypti cells,there were 20 mi RNAs expressed in dengue virus group alone and4 mi RNAs expressed differently in ZIKV virus group.In midgut tissues,6 mi RNAs were only expressed in the midgut of Ae.aegypti infected with DENV,14 mi RNAs were only expressed in the midgut of Ae.aegypti infected with ZIKV,35 mi RNAs were only expressed in the salivary glands of Ae.aegypti infected with DENV,and 30 mi RNAs were only expressed in the salivary glands of Ae.aegypti infected with ZIKV.The analysis of differential gene expression in midgut and salivary gland tissues of Ae.aegypti revealed that 285 genes were expressed in dengue virus group and 595 genes were expressed in ZIKV virus group in midgut tissues;891 genes were expressed in dengue virus group and 1620 genes were expressed in Zika virus group in salivary gland tissues.(4)Go and KEGG enrichment analysis of differentially expressed genes showed that the expression of differentially expressed genes mainly regulated cell metabolism,cell cycle,apoptosis and immune response,especially membrane composition and membrane function,such as cell membrane,endoplasmic reticulum membrane,Golgi membrane and so on.(5)The results of q RT-PCR were consistent with that of RNA-seq.It could be concluded:(1)After Ae.aegypti cells were infected with DENV-2and ZIKV,the speed of ZIKV replication is faster.(2)After Ae.aegypti cells were infected with DENV-2 and ZIKV,the incubation period of Ae.aegypti infected with ZIKV is shorter,and the infection rate and viral load of ZIKV were higher in all tissues of Ae.aegypti.(3)The m RNA and mi RNAs expression profiles of Ae.aegypti infected with DENV-2 and ZIKV were obtained and analyzed.(4)The accuracy of sequencing data was confirmed by QRT PCR of Ae.aegypti cells infected with DENV-2and ZIKV and the differentially expressed genes of Ae.aegypti.