O Método ACRS (Amplification-Created Restriction Site) na Detecção do Polimorfismo rs2227306 no Gene Interleucina (Il8) e Comparação Interétnica da Distribuição Genotípica

Abstract

The single nucleotide polymorphism (SNP) +781(C/T) (rs2227306) in the interleukin 8 gene (IL8) has been largely investigated in case-control association studies. Different methods have been used to genotype individuals for this SNP, but they demand extensive optimization of polymerase chain reaction (PCR) conditions or high cost reagents and equipment. The aim of this study was to develop a simple and efficient assay for the detection of the polymorphism rs2227306 in the IL8 gene and to execute an interethnic comparison of the genotype distribution in different populations. The Amplification-Created Restriction Site (ACRS) method was used for genotyping 174 healthy Brazilian individuals (Whites n=148; Blacks n=26). This method was efficient, reproducible and accurate, since the genotypes were confirmed by sequencing. White Brazilian subgroup showed a genotype distribution similar to HapMap CEU Europeans (p=0.49), but different from a German population, PGA African American and HapMap Sub-Saharan Africans (p<0.05). The genotype distribution in Black Brazilians was similar to that reported to PGA African American (p=0.635), but it was different from Sub-Saharan Africans, HapMap-CEU Europeans and Germans. Interethnic comparison demonstrated that Germans carried the highest T allele frequency (45.7%) and Sub-Saharan Africans carried the lowest T allele frequency (6.7%). The ethnically admixed Brazilian population showed intermediate frequencies of the T allele. We conclude that this ACRS method for genotyping the SNP rs2227306 in the IL8 gene was very accurate, simple and convenient for limited technology laboratories worldwide. We believe this technique is useful for genotyping in case-control association and population genetics studies.