title: Facile and phase-defined determination of HLA alleles with morpholino-functionalized nanoparticle probes.
abstract: A number of human leukocyte antigen (HLA) gene alleles have been found to be genetic risk markers for immunologically mediated drug hypersensitivity. Clinical adoption of HLA pharmacogenomics requires facile and accurate allele screening assays. As HLA genes are highly polymorphic, currently available methods are usually labor-intensive and liable to generate false positives. Herein we report a general strategy for screening HLA alleles with nanoparticle probes. Specific HLA alleles can be identified by gauging three to five sequence variants. Single-polymerase chain reaction (PCR) and dual-PCR methods have been proposed to achieve phase-defined determination of the sequence variants. Morpholino-functionalized gold nanoparticle probes allow for colorimetric and highly specific detection. Assays for HLA-B*58:01 and HLA-B*15:02 have been developed and validated with 49 selected human genomic DNA samples. The facile nanoparticle probe-based assays can be implemented easily in molecular diagnostic laboratories for accurate and cost-effective screening of HLA alleles.
authors.affiliation: Institute of Bioengineering and Nanotechnology, Singapore, Singapore.
authors.forename: Yanbing
authors.initials: Y
authors.lastname: Zu
authors.affiliation: Institute of Bioengineering and Nanotechnology, Singapore, Singapore.
authors.forename: Min-Han
authors.initials: MH
authors.lastname: Tan
authors.affiliation: Singapore Immunology Network, 8A Biomedical Grove, Immunos, Singapore, Singapore.
authors.forename: Ee Chee
authors.initials: EC
authors.lastname: Ren
authors.affiliation: Institute of Bioengineering and Nanotechnology, Singapore, Singapore. Electronic address: jyying@ibn.a-star.edu.sg.
authors.forename: Jackie Y
authors.initials: JY
authors.lastname: Ying
mesh_terms: D000483:Alleles; D001483:Base Sequence; D006680:HLA Antigens; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D060172:Morpholinos; D053758:Nanoparticles; D000071185:Pharmacogenomic Testing / Q000379:methods*; D016133:Polymerase Chain Reaction
chemical_list: D006680:HLA Antigens; D015235:HLA-B Antigens; D060172:Morpholinos
keywords: Colorimetric detection; HLA alleles; Morpholino-functionalized gold nanoparticles; Pharmacogenomics; Screening