别嘌醇引发严重皮肤不良反应标志基因HLA-B*5801的检测方法研究

Research on Detection Methods for HLA-B*5801: A Biomarker for Allopurinol Induced Serious Cutaneous Adverse Drug Reactions

目的建立准确、快速、经济的别嘌醇引发严重皮肤不良反应标志基因HLA-B*5801检测方法。方法收集2012年1月—2014年12月福建汉族人群中服用单一别嘌醇致严重皮肤不良反应患者血清样本14例,并随机抽取同期服用别嘌醇14 d未出现皮肤不良反应的患者30例。入组患者DNA分别用聚合酶链-顺序特异性引物反应法(PCR-SSP)、聚合酶链-限制性长度多态性法(PCR-RFLP)和聚合酶链-直接测序法(PCR-SBT)检测HLA-B*5801基因。对检测结果的准确性,方法的操作性及经济性等进行比较分析。结果 3种方法取得一致的检测结果,别嘌醇重症药疹患者100%(14/14)携带HLA-B*5801基因,而别嘌醇耐受患者23.33%(7/30)携带该基因(OR=90.87,敏感度=100%,专属性=76.67%)。结论PCR-SSP法和PCR-RFLP法均能筛检HLA-B*5801,都属于快速、经济,特异性高、操作简便、实验条件稳定的检测方法,非常适合于一般条件的临床推广使用,具有一定应用价值。

OBJECTIVE To establish an accurate, rapid and inexpensive method for the detection of HLA-B＊5801 which was a biomarker for allopurinol induced serious cutaneous adverse drug reactions（SCADRs）. METHODS Collected 14 Chinese patients with allopurinol-SCADRs and 30 randomly inpatients taking allopurinol more than 14 d with not cutaneous adverse reactions from 2012 January to 2014 December in Fujian Han population. Extracted DNA, then detected HLA-B＊5801 by PCR-sequence specific primers （PCR-SSP）, PCR-restriction fragment length polymorphism （PCR-RFLP） and polymerase chain reaction-sequence based typing（PCR-SBT）. Analyzed the methods of accuracy of results, operation and economy, etc. RESULTS The three methods got the same result. It is found that the allopurinol-SCADRs patients 100%（14/14） had HLA-B＊5801, whereas only 23.33% （7/30） in the allopurinol-tolerant patients（OR=90.87, sensitivity=100%, specificity=76.67%） CONCLUSION PCR-SSP and PCR-RFLP were rapid, inexpensive, high specificity, easy operation and stability assays which would be useful for pre-screening the subjects with HLA-B＊5801, very suitable for clinical use in general and have great clinical value.