title: Development of new IL28B genotyping method using Invader Plus assay.
abstract: IL28B polymorphism is associated with the response to pegylated interferon-α with ribavirin (PEG-IFN-α/RBV) treatment in chronic hepatitis C patients. As a genotyping assay for IL28B single nucleotide polymorphisms (SNPs) in clinical practice, the Invader Plus assay was developed. The accuracy, intra-assay, inter-assay precision, and the limit of detection of the Invader Plus assay were evaluated. Two SNPs (rs8099917 and rs12979860) associated with IL28B were genotyped by the Invader Plus and TaqMan assay in 512 Japanese patients. In comparison with direct sequencing, the Invader Plus assay showed 99% accuracy in rs8099917 and 100% accuracy in rs12979860. Intra-assay and inter-assay precision were sufficient to use in clinical practice and the detection limit was 1ngDNA/assay. Genotyping by rs8099917 showed that 361 (71%), 144 (28%) and seven (1%) of the patients were major homozygous, heterozygous and minor homozygous types, respectively. Five of the 512 cases (1%) had haplotype differences, but none showed differences between the two genotyping methods. For patients with HCV genotype 1, the prevalence of responders in the major homozygous type was 83.3%, and that of non-responders in the minor heterozygous/homozygous type was 72.5%. A convenient IL28B genotyping method using the Invader Plus assay could be useful to predict the treatment outcome in clinical practice.
authors.affiliation: Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Sciences, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Kani
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Kentaro
authors.initials: K
authors.lastname: Matsuura
authors.forename: Tsunamasa
authors.initials: T
authors.lastname: Watanabe
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yatsuhashi
authors.forename: Etsuro
authors.initials: E
authors.lastname: Orito
authors.forename: Ken
authors.initials: K
authors.lastname: Inose
authors.forename: Nao
authors.initials: N
authors.lastname: Motojuku
authors.forename: Yukio
authors.initials: Y
authors.lastname: Wakimoto
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizokami
mesh_terms: D000328:Adult; D000368:Aged; D044466:Asian People / Q000235:genetics; D005260:Female; D005821:Genetic Techniques; D005838:Genotype; D019698:Hepatitis C, Chronic / Q000188:drug therapy / Q000235:genetics*; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use; D007372:Interferons; D007378:Interleukins / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D011933:Reagent Kits, Diagnostic; D012254:Ribavirin / Q000627:therapeutic use; D000095022:Interferon Lambda
chemical_list: D016898:Interferon-alpha; D007372:Interferons; D007378:Interleukins; D011933:Reagent Kits, Diagnostic; D012254:Ribavirin; C000624353:interferon-lambda, human; D000095022:Interferon Lambda