title: Simple assay based on restriction fragment length polymorphism associated with IL28B in chronic hepatitis C patients.
abstract: OBJECTIVE
  Several studies recently revealed that single nucleotide polymorphisms (SNPs) in the interleukin28B (IL28B) region are associated with the response to pegylated interferon-alfa (PEG-IFN-alfa) and ribavirin (RBV) treatment among hepatitis C virus (HCV)-infected individuals of European, African and Asian ancestry. The purpose of the study was to establish methods for determining the SNP rs8099917 associated with IL28B, which might be useful for further research of the treatment of HCV.
  
  
  METHODS
  Blood samples obtained from 93 consecutive patients with chronic hepatitis C were examined. On the basis of the sequence data, a new simple genotyping assay based on a PCR-restriction fragment length polymorphism (RFLP) with two enzymes, BsrDI and Tsp45I, was developed.
  
  
  RESULTS
  The proportion of null virological responders in the combined TG/GG group was higher than that in the TT group (p = 0.015), suggesting that minor allele is one of the important factors playing crucial roles in IFN-resistance. Genotyping of rs8099917 by our new method showed results identical to PCR and sequence in 98.9% and 98.9% by BsrDI and Tsp45I, respectively. Using two enzymes, BsrDI and Tsp45I, it was possible to distinguish IL28B SNP rs8099917.
  
  
  CONCLUSIONS
  This simple method using RFLP will provide the framework for further studies of HCV.
authors.affiliation: Department of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Nakamoto
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kanda
authors.forename: Fumio
authors.initials: F
authors.lastname: Imazeki
authors.forename: Shuang
authors.initials: S
authors.lastname: Wu
authors.forename: Makoto
authors.initials: M
authors.lastname: Arai
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fujiwara
authors.forename: Osamu
authors.initials: O
authors.lastname: Yokosuka
mesh_terms: D000328:Adult; D000368:Aged; D004262:DNA Restriction Enzymes; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D019698:Hepatitis C, Chronic / Q000188:drug therapy*; D006801:Humans; D007372:Interferons; D007378:Interleukins / Q000235:genetics*; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D000095022:Interferon Lambda
chemical_list: D004262:DNA Restriction Enzymes; D007372:Interferons; D007378:Interleukins; C000624353:interferon-lambda, human; D000095022:Interferon Lambda