Development of a Real-time PCR Assay of HLA-B*15:02 Allele for Prevention of Carbamazepine Induced Steven Johnson Syndrome/ Toxic Epidermal Necrolysis in Neurological Institute of Thailand
Keywords:
HLA-B*15:02, Real-time PCR, Steven-Johnson Syndrome (SJS), Toxic epidermal necrolysis (TEN), CarbamazepineAbstract
Background: Steven Johnson Syndrome (SJS) and Toxic epidermal necrolysis (TEN) are severe adverse drug reactions (ADR) that increase the morbidity and mortality. Most are hospitalized for a long period of time. This causes high treatment costs and possibly requires continuous treatment after being discharged from the hospital. Therefore, HLA-B*15:02 detection before drug dispensing can help physicians in selecting therapeutic drugs to prevent ADR or drug allergies in patients. Objectives: The researcher aimed at developing a real-time PCR assay for detection of HLA-B*15:02. Methods: EDTA whole blood samples were collected from 76 patients during 2022-2023, extract for DNA and screened for HLA-B*15:02 gene using the real-time PCR with in-house designed primers and probes specific for HLA-B*15:02 gene according to the HLA databases. Then, we compared the result of our developed real-time PCR assay with Sanger gene sequencing for HLA-B*15:02 gene genotyping. Results: In this study we found 29 positive HLA-B*15:02 (38.15%) and 47 negative HLA-B*15:02 were samples (61.85%) The sensitivity and specificity of this developing method were 100% and 100%, respectively. All samples were compared with Sanger sequencing and all results showed similar to sanger sequencing. Conclusion: This study was successful developed assay for detecting HLA-B*15:02 allele to prevent CBZ-induced SJS/TEN.
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