title: Development and Validation of a Simple, Rapid and Inexpensive PCR-RFLP Method for Genotyping of Common IL28B Polymorphisms: A Useful Pharmacogenetic Tool for Prediction of Hepatitis C Treatment Response.
abstract: BACKGROUND
  In 2009, 3 genome-wide association studies implicated IL28B single-nucleotide polymorphisms (SNPs) as the strongest genetic pretreatment predictor of sustained virological response (SVR) in hepatitis C infection. Recently, the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) included IL28B testing in their guidelines.
  
  
  OBJECTIVE
  The main aim of this study was to develop and validate a simple, rapid, and inexpensive polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for genotyping of common IL28B polymorphisms (rs12979860 and rs8099917).
  
  
  METHODS
  Two methods were developed to genotype common IL28B polymorphisms: 1) PCR-sequencing as a reference method and 2) PCR-RFLP as a rapid and inexpensive method. Both polymorphisms were genotyped in 104 Iranian hepatitis C patients by both methods simultaneously. To validate the PCR-RFLP method, the PCR-RFLP genotyping results should be 100% concordant with the PCR-sequencing results.
  
  
  RESULTS
  Genotyping of rs12979860 and rs8099917 by PCR-RFLP was concordant with PCR-sequencing in 104 (100%) individuals. The analytical sensitivity and specificity of the PCR-RFLP method for genotyping of both SNPs are 100%. Among these 104 patients with chronic hepatitis C, the frequency of the rs12979860 CC, CT and TT genotypes were 40.4%, 47.1% and 12.5% and the frequency of the rs8099917 TT, GT and GG genotypes were 59.6%, 35.6% and 4.8%, respectively. Also, three IL28B haplotypes (rs12979860-rs8099917) were found among our patients including C-T, T-G and T-T with 63.9%, 22.6% and 13.5% frequency, respectively. C-G haplotype was absent in all of our patients.
  
  
  CONCLUSIONS
  We have developed a validated, fast, and simple PCR-RFLP method for genotyping of common IL28B SNPs that is more cost-effective than sequencing.
authors.affiliation: Armin Pathobiology Laboratory, Tehran, IR Iran.
authors.forename: Heidar
authors.initials: H
authors.lastname: Sharafi
authors.forename: Ali
authors.initials: A
authors.lastname: Pouryasin
authors.forename: Seyed Moayed
authors.initials: SM
authors.lastname: Alavian
authors.forename: Bita
authors.initials: B
authors.lastname: Behnava
authors.forename: Maryam
authors.initials: M
authors.lastname: Keshvari
authors.forename: Leila
authors.initials: L
authors.lastname: Mehrnoush
authors.forename: Shima
authors.initials: S
authors.lastname: Salimi
authors.forename: Osveh
authors.initials: O
authors.lastname: Kheradvar