Copy Number Variation and gene Rearrangements in CYP2D6 Genotyping using Multiplex Ligation-dependent Probe Amplification in Koreans

Abstract

Aim: The present study introduces a simple method for CYP2D6 genotyping that not only determines the heterozygous or homozygous deletions and duplications, but also distinguishes tandem hybrids. Materials & methods: Using two commercially available methods, 49 Korean male subjects were genotyped for CYP2D6. The Affymetrix^® Targeted Human Drug Metabolizing Enzymes and Transporter 1.0 Assay was used for SNP genotyping and multiplex ligation-dependent probe amplification (MLPA) assay (SALSA^® MLPA^® Kit P128-A1 CYP450) was used for copy number analysis. Long range PCR was used to confirm the MLPA results. Fifty Caucasian samples obtained from the Coriell Institute were used to confirm the accuracy of the MLPA assay. Results: Using two commercially available methods, we found seven different allele types with CYP2D6*1 (34.7%), *2 (12.2%), *10 (17.4%) and *36-*10 (22.5%) being the most common alleles in the Korean population. The MLPA results showed 100% agreement with long-range-PCR results and were able to distinguish deletions and duplications among 50 Caucasian reference samples. Conclusion: The application of MLPA-based genotyping to routine clinical analysis will enable patients to be assigned to more accurate genotypes at a reasonable cost in a large number of individuals at the majority of locations.

KEYWORDS::

• CYP2D6 gene
• genotyping
• multiplex ligation-dependent probe amplification

Financial & competing interests disclosure

This work was supported by a grant from the Korea Healthcare technology R&D Project, Ministry of Health, Welfare & Family Affairs, Republic of Korea [A070001] and the faculty research grant of Yonsei University College of Medicine for 2011 (6-2011-0076). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

Financial & competing interests disclosure This work was supported by a grant from the Korea Healthcare technology R&D Project, Ministry of Health, Welfare & Family Affairs, Republic of Korea [A070001] and the faculty research grant of Yonsei University College of Medicine for 2011 (6-2011-0076). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.