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

  • Aline Cavalcanti Viana Faculdade de Odontologia de Araraquara - FOAr/UNESP
  • Karen Maria de Carvalho Curtis Faculdade de Odontologia de Araraquara - FOAr/UNESP
  • Yeon Jung Kim Faculdade de Odontologia de Araraquara - FOAr/UNESP
  • Silvana Regina Perez Orrico Faculdade de Odontologia de Araraquara - FOAr/UNESP
  • Veridiana S.P. Cano Faculdade de Ciências Farmacêuticas de Araraquara - FCFAr/UNESP
  • Sandro Roberto Valentini Faculdade de Ciências Farmacêuticas de Araraquara - FCFAr/UNESP
  • Raquel Mantuaneli Scarel-Caminaga Faculdade de Odontologia de Araraquara - UNESP

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.

Author Biographies

Aline Cavalcanti Viana, Faculdade de Odontologia de Araraquara - FOAr/UNESP
Dentist; Msc. in Periodontology; Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP - São Paulo State University, Araraquara, SP, Brazil. E-mail: dentistaaline@yahoo.com
Karen Maria de Carvalho Curtis, Faculdade de Odontologia de Araraquara - FOAr/UNESP
Biologist; Department of Morphology, School of Dentistry at Araraquara, UNESP - São Paulo State University, Araraquara, SP, Brazil. E-mail: karcurtis@iq.unesp.br
Yeon Jung Kim, Faculdade de Odontologia de Araraquara - FOAr/UNESP
Dentist; Msc in Periodontology; Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP - São Paulo State University, Araraquara, SP, Brazil. E-mail: yeon_jungkim@yahoo.com.br
Silvana Regina Perez Orrico, Faculdade de Odontologia de Araraquara - FOAr/UNESP
Dentist; PhD in Periodontology; Professor, Department of Oral Diagnosis and Surgery, School of Dentistry at Araraquara, UNESP - São Paulo State University, Araraquara, SP, Brazil. E-mail: s_orrico@foar.unesp.br
Veridiana S.P. Cano, Faculdade de Ciências Farmacêuticas de Araraquara - FCFAr/UNESP
Pharmaceutic; PhD in Biotechnology, Department of Biological Sciences, School of Pharmaceutical Sciences, UNESP – São Paulo State University, Araraquara, SP, Brazil. E-mail: canovsp@fcfar.unesp.br
Sandro Roberto Valentini, Faculdade de Ciências Farmacêuticas de Araraquara - FCFAr/UNESP
Pharmaceutic; PhD in Biochemistry; Professor, Department of Biological Sciences, School of Pharmaceutical Sciences, UNESP – São Paulo State University, Araraquara, SP, Brazil. E-mail: valentsr@fcfar.unesp.br
Raquel Mantuaneli Scarel-Caminaga, Faculdade de Odontologia de Araraquara - UNESP
Biologist; PhD in Buco-Dental Biology, Professor, Department of Morphology, School of Dentistry at Araraquara, UNESP - São Paulo State University, Araraquara, SP, Brazil. E-mail: raquel@foar.unesp.br
Published
2009-06-08
Section
Artigos Originais