Facile and phase-defined determination of HLA alleles with morpholino-functionalized nanoparticle probes

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.

Graphical AbstractScreening of specific human leukocyte antigen (HLA) gene alleles, such as HLA-B*15:02 and HLA-B*58:01, has been adopted in clinical practice to reduce the risk of drug hypersensitivity. However, the highly polymorphic nature of HLA genes makes it difficult to achieve reliable tests with simple protocols. Herein we report a general strategy as well as two assays as examples for rapid screening of HLA alleles with morpholino-functionalized nanoparticle probes. The nanoparticle probes allow for colorimetric and highly specific detection. The facile and phase-defined assays can be implemented easily in molecular diagnostic laboratories.207