Development of a Simple Genotyping Method for the Hla-A*31:01-Tagging Snp in Japanese

Abstract

Aim: To construct a simple, low-cost typing method for the surrogate marker of HLA-A*31:01, a risk factor for carbamazepine (CBZ) related Stevens–Johnson syndrome/toxic epidermal necrolysis (SJS/TEN). Materials & methods: DNAs from Japanese SJS/TEN patients were used for genotyping and developing the assay. Results:HLA-A*31:01 was confirmed to be significantly associated with definite/probable cases of CBZ-related SJS/TEN (p = 0.0040). Three single nucleotide polymorphisms, rs1150738, rs3869066 and rs259945, were in absolute linkage disequilibrium with HLA-A*31:01 in 210 Japanese SJS/TEN patients. Robust genotyping of rs3869066 in ZNRD1-AS1 was developed using polymerase chain reaction-restriction fragment length polymorphism assays. Conclusion: Single nucleotide polymorphism genotyping is less time consuming and cheaper than conventional HLA typing, and would be useful for identifying Japanese patients at risk of CBZ-related SJS/TEN.

Keywords::

• carbamazepine
• PCR-RFLP
• screening test
• Stevens–Johnson syndrome/toxic epidermal necrolysis

Acknowledgements

The authors thank all the patients and physicians for their co-operation. The authors deeply appreciate the assistance of the Ministry of Health and Labour and Welfare (MHLW), Pharmaceuticals and Medical Device Agency (PMDA), the Federation of Pharmaceutical Manufacturers’ Association of Japan (FPMAJ) in recruiting patients.

Financial & competing interests disclosure

This study was supported in part by the Health Labour Sciences Research Grants from the Ministry of Health, Labour, and Welfare, Japan Agency for Medical Research and development, AMED and the JSPS KAKENHI. 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

This study was supported in part by the Health Labour Sciences Research Grants from the Ministry of Health, Labour, and Welfare, Japan Agency for Medical Research and development, AMED and the JSPS KAKENHI. 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.