Validation of a novel real-time PCR assay for detection of HLA-B*15:02 allele for prevention of carbamazepine - Induced Stevens-Johnson syndrome/Toxic Epidermal Necrolysis in individuals of Asian ancestry
Validation of a novel real-time PCR assay for detection of HLA-B*15:02 allele for prevention of carbamazepine - Induced Stevens-Johnson syndrome/Toxic Epidermal Necrolysis in individuals of Asian ancestry
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DOI：
10.1016/j.humimm.2016.08.010
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发表时间：
2016-12-01
期刊：
影响因子：
2.7
通讯作者：
Fernando, Suran L.
中科院分区：
文献类型：
--
作者：
Dinh Van Nguyen;Vidal, Christopher;Fernando, Suran L.
目的：建议筛查HLA-B*15：02等位基因，以预防亚裔个体中卡马西平（CBZ）诱导的史蒂文斯·约翰逊综合征（SJS）和中毒性表皮坏死松解症（TEN）。因此，我们的目的是开发和验证一种稳健和廉价的方法来检测HLA-B*15：02等位基因。方法：使用TaqMan（R）探针和SYBR（R）绿色的实时PCR在CBZ治疗前检测HLA-B*15：02等位基因。结果：共检测了121个样本。该测定法的灵敏度为100%（95% CI：76.84-100.0%），特异性为100%（95% CI：96.61-100%），阳性预测值为100%（95% CI：76.84-100%），阴性预测值为100.0%（95% CI：96.61-100.0%）。我们的结果与使用Luminex SSO/SBT/SSP进行的基因分型结果100%一致。TaqMan探针的最低检测限为0.05 ng/μ l，SYBR绿色为0.5 ng/μ l DNA。使用TaqMan（R）探针和SYBR绿色探针的单位成本仅为4.7美元。结论：我们开发了一种检测HLA-B*15：02等位基因的新方法，该方法稳定、廉价，适用于筛查亚洲血统的个体，以预防CBZ诱导的SJS/TEN。(C)2016年美国组织相容性和免疫遗传学学会。爱思唯尔公司出版All rights reserved.
Objectives: Screening for the HLA-B*15:02 allele has been recommended to prevent carbamazepine (CBZ) - induced Stevens Johnson syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) in individuals with Asian ancestry. We aimed, therefore, to develop and validate a robust and inexpensive method for detection of the HLA-B*15:02 allele.Methods: Real-time PCR using TaqMan (R) probes followed by SYBR (R) Green was used to detect the HLA-B*15:02 allele prior to treatment with CBZ therapy.Results: A total of 121 samples were tested. The assay has a sensitivity of 100% (95% CI: 76.84-100.0%), a specificity of 100% (95% CI: 96.61-100%), a positive predictive value of 100% (95% CI: 76.84-100%) and a negative predictive value of 100.0% (95% CI: 96.61-100.0%), respectively. There was 100% agreement between our results and genotyping using Luminex SSO/SBT/SSP. The lowest limit of detection of the TaqMan (R) probe is 0.05 ng/mu l and the SYBR Green is 0.5 ng/mu l of DNA. The unit cost of using the TaqMan (R) probe followed by SYBR Green is only $4.7 USD.Conclusion: We developed a novel assay for the detection of the HLA-B*15:02 allele, which is robust, inexpensive and suitable for screening individuals of Asian ancestry in the prevention of CBZ-induced SJS/TEN. (C) 2016 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.