Development in detection of hla-B*1502 and –B*5801 by ssp-pcr and lamp with pna probe

Abstract

Pharmacogenetics in HLA allele is very useful for risk assessment in life-threatening drug hypersensitivity. In 2004, HLA-B*1502 first showed strong association with carbamazepine hypersensitivity. Another association, HLA-B*5801 involved with allopurinol hypersensitivity. Since allele frequencies of these HLA-B alleles are quite common in Asian ethnicity and with 26% mortality rate in patients, it is important to develop a rapid and cost effective test for Asian, including our Thai population. To get a HLA type, commercially available SSP-PCR is too expensive. So, in-house SSP-PCR would be the most interesting candidates comparing to other molecular techniques. From this study, HLA-B*1502 can be interpreted specifically by simple nested SSP-PCR, while HLA-B*5801 can be differentiated by only one set of primer with 100% sensitivity and >99.9% specificity. Moreover, blood PCR kit can be applied to both HLA-B*1502 and HLA-B*5801 with the same specificity. Furthermore, LAMP was introduced to HLA typing as an alternative method. LAMP is an isothermal amplification technique which its specificity comes from 4 primers at a time. In this study, HLA-B*5801 can be identified with 95.24% sensitivity and 83.78% specificity. The benefit of these tests would help patients to avoid any life-threatening adverse consequences.