HLA-B^(*)15∶02基因型的快速鉴别方法建立及临床应用  

Establishment and clinical application of a rapid identification method for HLA-B^(*)15∶02 genotype

作　　者：初亚男 封利颖 李玉娇 张婕妤 CHU Ya′nan;FENG Liying;LI Yujiao;ZHANG Jieyu(Department of Clinical Pharmacy,Eastern Theater Command General Hospital of PLA,Nanjing,Jiangsu 210002,China)

机构地区：[1]中国人民解放军东部战区总医院临床药学科,江苏南京210002

出　　处：《国际检验医学杂志》2023年第9期1031-1034,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金项目(82001422);中国人民解放军东部战区总医院院内课题(22LCZLXJS56)。

摘　　要：目的建立rs144012689位点的焦磷酸测序方法,从多个角度评估其预测芳香族类抗癫痫药物严重皮肤不良反应的性能。方法筛选最优的聚合酶链反应(PCR)扩增体系,评估该研究建立的rs144012689位点焦磷酸测序方法的检测限、灵敏度、特异度,并分析其与Sanger测序、商品化试剂盒的一致性。收集该院2019-2021年确诊为史蒂文斯-约翰逊综合征(SJS)的患者,评价该方法对确诊病例的检出性能。结果该方法检测rs144012689位点筛选出人类白细胞抗原(HLA)-B 15∶02基因型的灵敏度和特异度均为100%,检测限为0.4 ng/μL基因组DNA。60份标本焦磷酸测序结果均与Sanger测序及商品化试剂盒检测结果一致。2份SJS确诊患者标本的焦磷酸测序和HLA-B高分辨分型结果均提示携带HLA-B 15∶02基因型。结论该研究建立的方法具备快速、检测限低、灵敏度高和特异度高的优点,可用于芳香族类抗癫痫药物不良反应高危基因HLA-B 15∶02的筛查。Objective To establish a pyrosequencing method of rs144012689 site and to evaluate its efficacy in predicting severe skin antiepileptic adverse drug reaction in aromatic family from multiple perspectives.Methods The optimal PCR amplification system was screened and established.The detection limits,sensitivity,and specificity of the pyrosequencing method at rs144012689 site established in this study were evaluated.Meanwhile,this method was validated with Sanger sequencing and commercial kit.Patients diagnosed with Stevens-Johnson syndrome(SJS)from 2019 to 2021 in this hospital were collected to evaluate the detection performance of this method.Results The sensitivity and specificity of the method for detecting human leukocyte antigen(HLA)-B 15∶02 genotype at rs144012689 were 100%and the detection limit was 0.4 ng/μL genomic DNA.The pyrosequencing results of 60 samples were consistent with those of Sanger sequencing and commercial kit.Pyrosequencing and HLA-B high resolution typing results of 2 samples of confirmed SJS patients indicated that they carried HLA-B 15∶02 genotype.Conclusion The method established in this study has the advantages of rapid detection,low detection limit,high sensitivity and high specificity,and it could be used for the screening of high risk gene HLA-B 15∶02 of antiepileptic adverse drug reaction in aromatic family.

关 键 词：rs144012689位点 抗癫痫药物 HLA-B 15∶02 焦磷酸测序 基因型 史蒂文斯-约翰逊综合征 

分 类 号：R446.9[医药卫生—诊断学]