Establishment Of Real-time Quantitative PCR And Bioinformatics Analysis Of African Genetype Zika Virus Base On E Gene

Zika virus is a mosquito-borne flavivirus(a member of the family Flaviviridae)related to yellow fever virus,dengue virus,and West Nile virus.Nowadays,ZIKV isolates can be grouped into two major lineages,These lineages correspond to the African lineage,the Asian lineage.In brief,Zika virus(ZIKV)was first isolated in 1947 from a febrile sentinel rhesus monkey held in a cage on a platform in the canopy of the Zika Forest in Uganda during studies to identify the vector of sylvatic YF,For the next few decades,ZIKV has been a "sleep state",causing scattered transmission,and the symptoms are lighter and self-healing.Brazil reported the first cases of locally transmitted ZVI in the Americas,and spread to other parts of the world quickly.The increase in GBS cases and microcephaly cases,a surge in the number of patients,an expanded range of transmission set off international alarms and garnered global attention,and ZIKV infection is a serious threat to human health.Zika virus since its declaration as a public health emergency of international concern in 2016.Sequence homology studies reveal that of the two ZIKV lineages,the strains responsible for the human outbreaks were phylogenetically closest to the Asian lineageThe study established a high specificity,sensitivity and repeatability African genetype Zika virus detection method base on African standard strain MR766,which can provide support for prevention and control of African genetype Zika virus.The standard curve of the standard recombinant plasmid has a good linear relationship between 2.47×108?102copies/?l.This method can specifically detect the standard plasmid without cross reactions with Dengue type ???,Asian genetype Zika virus,HCV-1b RNA,JEV and CHIKV.The detection limit of this method was 2.47×101copies/?l.Therefore,the study can be applied to the detection of clinical suspicious specimens,and can realize early rapid detection,differential diagnosis and virus quantification in African ZIKV.ZIKV E protein mediates the entry of the virus into the host cell by binding to the host cell surface receptor to initiate the infection cycle of ZIKV.It is a key factor in initiating ZIKV entry into the host cell and participates in the whole process of the viral cycle.By analyzing the mutation direction of the amino acid sequences of the two lineage E proteins,it was found that the mutation direction of the amino acid site was closely related to the evolution of the African ZIKV to the Asian ZIKV and caused the outbreak infection,and the amino acid mutation also caused the ZIKV epitope.change.This study mainly analyzes the E protein bioinformation,infection source and transmission route,historical and geographical distribution,clinical manifestation and detection technology status of Zika virus,and establishes the rapid detection technology of African ZIKV to provide Zika virus disease.Rapid diagnostic techniques provide a strong basis for subsequent treatment;and bioinformatics analysis of the E protein of the two lineages of ZIKV is carried out,hoping to provide a useful reference for further research of ZIKV.