Detection of the single nucleotide polymorphism (rs2227307) in the human interleukin 8 gene using a different PCR-RFLP assay

Abstract

Interleukin 8 (IL-8) is chemokine that acts as a potent chemoattractant for neutrophils. Single nucleotide polymorphisms (SNPs) in the human IL8 gene have been investigated in many studies for disease association. We have developed a different PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment of Length Polymorphism) assay for genotyping the SNP (rs2227307) in the IL8 gene. This method was utilized in order to typing 147 white healthy Brazilian individuals whose DNA were obtained from epithelial buccal cells and extracted with phenol:chlorophorm:isoamilic alcohol. Genomic DNA was amplified by PCR using a conventional thermal cycler; following the PCR products (573 bp) were submitted to the RFLP reactions. The RFLP products were analysed in a 4% agarose gel stained with ethidium bromide. The genotype distribution observed in this study was consistent with the assumption of Hardy-Weinberg equilibrium and was similar (p=0.30) to those reported for other white populations at the website of the National Center for Biotechnology Information (NCBI) SNP Database. Because the PCR-RFLP method presented here was efficient, low cost, reproducible and convenient for worldwide technology limited laboratories, should be useful for genotyping in case-control association or population genetic studies.