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ORIGINAL ARTICLE
Year : 2016  |  Volume : 3  |  Issue : 3  |  Page : 73-76

Different methodological approaches for interleukin 28B genotyping


Department of Clinical Biochemistry, Molecular Biology Laboratory, Facultad de Química, Universidad de la República, Montevideo, Uruguay

Correspondence Address:
Dr. Patricia Esperón
General Flores 2124, CP 11800. Montevideo
Uruguay
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2225-6482.191368

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Background and Objectives: Treatment responsiveness to pegylated interferon-α and ribavirin against hepatitis C virus genotype 1 has strongly been associated with two single nucleotide polymorphisms (rs8099917 and rs12979860) in the region of the interleukin 28B (IL28B) gene. The aim was to perform three genotyping methods and evaluate their specificity, technical characteristics, and costs. In addition, the distribution of both polymorphisms in an Uruguayan population was assessed. Materials and Methods: One hundred DNA samples were genotyped by allele-specific polymerase chain reaction (AS-PCR), real-time PCR high resolution melting (RT-HRM), and Sanger sequencing methods. Results: The rs12979860 CC genotype, followed by the CT, was the most prevalent (52% and 39%, respectively). For rs8099917, the TT genotype was the most common (61%) followed by the GT (34%). AS-PCR and RT-HRM assays were specific for both IL28B genotypes determinations and showed a total concordance with Sanger sequencing results. Conclusions: Any of three genotyping methods is suitable for IL28B genotyping. The choice of the assay will depend on costs, special equipment availability, turnaround time, and specialized human resources.


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