Mutation screening of 3’UTR and exons 1-2 of vsx1 gene by PCR-SSCP/HA and sequencing in patients with Vernal Keratoconjuctivis (VKC) in Shahrekord

Abstract

Background and Objective: Vernal Keratoconjuctivis is an immune response in relation to environmental antigens, leading to inflammation of the conjunctiva. One of the presumable genetic factors in VKC is VSX1 gene. In this study, mutations in exon 1, exon 2 and 3’UTR of VSX1 gene in patients with VKC in Shahrekord were investigated by PCR-SSCP and PCR-HA. Materials and Methods: In this cross- sectional study, peripheral blood samples of 100 patients with VKC and 100 individuals with no confirmed eye disease as control group were investigated. Genomic DNA was extracted by phenol-chloroform method and then PCR was carried out. Then, SSCP and HA were performed and the samples with shifted bands were sequenced for the type of nucleotide change. Afterwards, to investigate the observed nucleotide change, RFLP method was used. Results: Our SSCP findings revealed six patients with shifted band in exons 1 and 2 and 13 patients in 3’UTR, which were sequenced for nucleotide change. Analysis of sequencing data showed a frameshift change (g. 25057561delG) in 3’UTR. There was no change in other sequences. Conclusion: The findings of this study showed that, VSX1 gene most probably has no effective role in VKC pathogenesis in the studied population. Therefore, the role of VSX1 genes in VKC pathogens needs further investigation. © 2015 Zanjan University of Medical Sciences and Health Services.