Multiplex SNaPshot-a new simple and efficient CYP2D6 and ADRB1 genotyping method

作者：Ben Songtao; Cooper DeHoff Rhonda M; Flaten Hanna K; Evero Oghenero; Ferrara Tracey M; Spritz Richard A; Monte Andrew A^*

来源：Human Genomics, 2016, 10(1): 11.

DOI：10.1186/s40246-016-0073-3

• ADRB1
• CYP2D6
• Copy number variation
• Gene deletion
• Genotyping
• Personalized medicine
• Precision medicine

Background: Reliable, inexpensive, high-throughput genotyping methods are required for clinical trials. Traditional assays require numerous enzyme digestions or are too expensive for large sample volumes. Our objective was to develop an inexpensive, efficient, and reliable assay for CYP2D6 and ADRB1 accounting for numerous polymorphisms including gene duplications.
Materials and methods: We utilized the multiplex SNaPshot (R) custom genotype method to genotype CYP2D6 and ADRB1. We compared the method to reference standards genotyped using the Taqman Copy Number Variant Assay followed by pyrosequencing quantification and determined assigned genotype concordance.
Results: We genotyped 119 subjects. Seven (5.9 %) were found to be CYP2D6 poor metabolizers (PMs), 18 (15.1 %) intermediate metabolizers (IMs), 89 (74.8 %) extensive metabolizers (EMs), and 5 (4.2 %) ultra-rapid metabolizers (UMs). We genotyped two variants in the beta 1-adrenoreceptor, rs1801253 (Gly389Arg) and rs1801252 (Ser49Gly). The Gly389Arg genotype is Gly/Gly 18 (15.1 %), Gly/Arg 58 (48.7 %), and Arg/Arg 43 (36.1 %). The Ser49Gly genotype is Ser/Ser 82 (68.9 %), Ser/Gly 32 (26.9), and Gly/Gly 5 (4.2 %). The multiplex SNaPshot method was concordant with genotypes in reference samples.
Conclusions: The multiplex SNaPshot method allows for specific and accurate detection of CYP2D6 genotypes and ADRB1 genotypes and haplotypes. This platform is simple and efficient and suited for high throughput.

• 出版日期2016-4-23