Accurate identification of HLA-B*15:02 allele by two-dimensional polymerase chain reaction

Abstract

Background: HLA-B*15:02 is highly associated with carbamazepine-induced SJS/TEN; however, there is no rapid and accurate detecting method. Here, we present a method to distinguish HLA-B*15:02 from 16 highly homologous HLA-B*15 alleles.

Methods: The high-throughput two-dimensional polymerase chain reaction (2D-PCR) technology was employed to identify HLA-B*15:02 in two-tube reaction. And, 2D-PCR accuracy was verified by PCR-sequence based typing (PCR-SBT).

Results: HLA-B*15:02 heterozygotes were identified by 14 melting valleys in the first tube reaction and none in the second, or by 13 melting valleys in the first tube reaction and one in the second. HLA-B*15:02 homozygote was identified by 13 melting valleys in the first tube reaction and none in the second. Three (0.16%) HLA-B*15:02 homozygotes and 84 (4.59%) HLA-B*15:02 heterozygotes were detected in 1830 samples of clinical general population without detecting 16 highly homologous alleles to HLA-B*15:02. The kappa test showed 100% coincidence between the 2D-PCR and PCR-SBT.

Conclusions: 2D-PCR in two-tube reaction method for identifying HLA-B*15:02 was successfully established. Identification of HLA-B*15:02 is necessary prior to taking CBZ based on HLA-B*15:02 allele frequency.

Keywords: 2D-PCR; Genotyping; HLA-B*15:02.