Phylogenetic Analysis of Dengue Virus Based on CprM Sequencing

Abstract

Dengue virus,a member of Flavirus causes an arboviral disease known as dengue fever. Dengue fever is an ailment that is transmitted by mosquitoes which is known as Aedes aegpti and up to some extent it is also spread by the Aedes albopictus. It is classified into distinct serotypes, mainly DENV i i i i i-1, and DENV-2, DENV 3, DENV-4, and DENV-5. More than half of the world's population are at high risk of DENV infection. In addition,up to 30–fold increase in dengue infection has been reported in the previous 50 years.The existing study was aimed to explore the CprM region of dengue virus in dengue positive isolates for nucleotide sequencing and its phylogenetic analysis in Pakistani population. In our study, blood samples from dengue infected patients were collected from Islamabad Pakistan. The samples were processed for RNA extraction followed by complementary DNA synthesis. Using high fidelity DNA polymerase, the CprM gene was amplified through nested PCR and the amplified product was further processed for nucleotide sequencing. In addition to the viral isolates from the current study, 68 refrence nucleotide sequences included in the study. Phylogenetic analysis of the current study sequence isolates (QAU IF- 1) (QAU-IF- 2) suggests clustering with a previously reported sequence from India with accession number (accession No. KU948528,KU948530).Further studies are required to explore amino-acid substitutions in functionally and structurally important regions of CprM protiens.