[Cloning and evaluation of expression of a novel overlapping region of NS3 gene of Hepatitis C virus by expressing vector]

In this project, our aim was to construct a novel expressing vector harboring a new sequence from overlapping region of NS3 gene of HCV from infected Iranian patient. The partial NS3 [pNS3] gene was amplified by Nested-RT-PCR method using sera of HCV infected patients harboring genotype 1a. After purification and cloning the pNS3 into TA-cloning vector, the best colony was selected based on Blue/White screening and colony-PCR following by confirmation with sequencing and restriction digestion with BglII. The sequenced gene was compared with other reference sequences using alignment softwares. The resultant pNS3 gene subcloned into the expression vector, IRES vector, followed by selection the suitable clones by 2 different colony-PCRs. The gene expression was evaluated using GFP detection, RT-PCR and western blotting techniques after transfection of the IRES-pNS3 vector into the 293 cell line. After pNS3 sequence amplification by RT-PCR, sequencing results showed high homology among the sequences with other reference sequences. This result also showed that it belonged to genotype 1 of HCV. Colony-PCR showed the insertion of gene into expressing vector with the right orientation. GFP expression, RT-PCR and western blotting confirmed transfection of vector, expression of pNS3 gene and production of its protein in 293 cells respectively. This novel expressing vector harboring partial region of NS3 gene in compare to full NS3 gene maybe more useful in immune induction by antigen presenting cells due to absence of genes responsible for protease activity of the protein in the setting of HCV vaccine

Seroprevalence of Varicella Zoster Virus in Children from Shiraz-Iran

Varicella-zoster virus [VZV] causes herpes zoster and varicella [Chicken-pox], usually a mild disease which is diagnosed clinically with few complications. However, in neonates and healthy adults it can have a severe presentation. Herpes zoster results from VZV reactivation later in life. To determine the seroprevalence of VZV in elementary school children aged 6-10 years in Shiraz, Iran. A cross-sectional seroprevalence survey was conducted on 270 healthy subjects. All serum samples were investigated for immunoglobulin G [IgG] antibody against VZV using a commercial enzyme linked immunosorbent assay [ELISA]. Among the studied population, 175 [64.8%] had no detectable antibody levels. The overall seroprevalence rate was 35.2%. A breakdown of seropositivity to VZV according to age was as follows; 10 years old, 50%, 9 years old, 48.2%, 8 years old, 27.3%, 7 years old, 32.1%, and 6 years old, 13.2%. As VZV susceptibility in the studied age groups was higher than the expected rate, therefore childhood VZV vaccination is recommended in our region