多色探针熔解曲线法在卡马西平不良反应相关基因检测中的临床评价

目的 对多色探针熔解曲线法（multicolor melting curve analysis，MMCA）用于卡马西平不良反应相关的HLA-B^*15：02基因检测进行临床评价。方法 收集厦门市中心血站1 147份厦门地区无偿献血者的外周静脉血样本，经基因DNA提取后，按双盲对照试验，分别应用MMCA法和HLA-SBT测序法对各样本进行HLA-B^*15：02基因检测，比较2种检测方法的符合率。对于检测结果不一致的标本，采用第三方Sanger测序和电泳验证，计算总符合率。结果 采用MMCA法共检出77份HLA-B^*15：02阳性标本，1 070份HLA-B^*15：02阴性标本。采用HLA-SBT测序法共检出74份HLA-B^*15：02阳性标本，1 070份HLA-B^*15：02阴性标本，以及3份无明确的分型信息的标本（仅显示为B^*15：VG-B^*15：CYS型）。该3份标本经Sanger测序以及电泳验证，确认为HLA-B^*15：02阳性标本。因此，MMCA法检测HLA-B^*15：02基因的阳性检出率为100%（77/77），阴性检出率为100%（1 070/1 070），总符合率为100%（1 147/1 147）。此外，在1 147份临床标本中共检出77份阳性结果，HLA-B^*15：02的携带率为6.7%（77/1147），这与文献报道的数据基本一致。结论 MMCA法用于HLA-B^*15：02基因的检测，具有简便、快速、灵敏度高、特异性强等优点，可应用于卡马西平不良反应相关的HLA-B^*15：02基因的临床辅助检测。

Clinical Evaluation of Multicolor Melting Curve Analysis for Detecting Gene Related to Carbamazepine-induced Adverse Drug Reactions

OBJECTIVE To evaluate the effect of multicolor melting curve analysis (MMCA) for detecting HLA-B^*15:02 gene related to carbamazepine-induced adverse drug reactions.METHODS A blind test of 1 147 peripheral blood samples collected from Xiamen Blood Center was performed to validate the agreement between MMCA assay and HLA-SBT assay. Sanger sequencing and electrophoresis were used to confirm inconsistent results of two methods,the agreement for all methods was validated.RESULTS In this study,77 cases were identified as positive samples and 1 070 cases were identified as negative samples by MMCA assay.For HLA-SBT assay,74 HLA-B^*15:02 positive samples,1 070 negative samples and 3 uncertain samples (only showed B^*15:VG-B^*15:CYS) were detected.Three uncertain samples were confiremed to be HLA-B^*15:02 positive samples by Sanger sequencing and electrophoresis.Therefore,the positive agreement,negative agreement and total agreement of HLA-B^*15:02 gene detection by MMCA assay were 100%(77/77),100%(1 070/1 070) and 100% (1 147/1 147).The percentage of positive samples was 6.7%(77/1 147) which was reasonable compared to previous reports. CONCLUSION MMCA assay is a convenient and rapid method with high sensitivity and excellent specificity,and promises to be a powerful tool for the clinical diagnosis of HLA-B^*15:02 gene.