title: A gene-specific primer extension and liquid bead array system for killer-cell immunoglobulin-like receptor genotyping.
abstract: A simple and accurate method for killer-cell immunoglobulin-like receptor (KIR) genotyping is developed using KIR gene-specific primer extension (GSPE) followed by bead array hybridization (GSPE method). After amplification of exons 4, 5, and 9, KIR GSPE and bead array hybridization were performed to verify the presence or absence of 16 KIR subfamilies. GSPE method was validated with natural killer/KIR reference panel I consisting of 48 cell types provided by 13th International Histocompatibility Working Group (IHWG) and genomic DNA from 17 peripheral blood cells, 8 cell lines, and 8 buccal cells. The results of reference panel from GSPE method were 100% concordant with the IHWG reference typing information. All genomic DNAs except reference panel were typed for KIR genes with sequence-specific primer methods and showed 100% identical typing results using this novel system. In addition, GSPE method can obtain results in 8 h from DNA with 10 ng genomic DNA in a 96-well-based assay format.
authors.affiliation: YeBT Co., Ltd, Mok-dong, Yangchun-gu, Seoul, South Korea.
authors.forename: H J
authors.initials: HJ
authors.lastname: Park
authors.forename: Y
authors.initials: Y
authors.lastname: Oh
authors.forename: H J
authors.initials: HJ
authors.lastname: Kang
authors.forename: E J
authors.initials: EJ
authors.lastname: Han
authors.forename: H Y
authors.initials: HY
authors.lastname: Shin
authors.forename: H S
authors.initials: HS
authors.lastname: Ahn
authors.forename: K S
authors.initials: KS
authors.lastname: Ahn
authors.forename: B H
authors.initials: BH
authors.lastname: Yoon
authors.forename: B D
authors.initials: BD
authors.lastname: Han
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics*; D005838:Genotype; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods; D008863:Microspheres; D008954:Models, Biological; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D054340:Receptors, KIR / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; D054340:Receptors, KIR