【摘要】 目的:通过检测评估荧光定量聚合酶链反应(PCR)染料法对人类白细胞抗原B位点5801(HLA-B*5801)等位基因的准确性、灵敏度及其在干扰因素下的结果分析研究，同时与PCR-SBT测序法进行对比。方法:收集45例痛风、高尿酸血症患者和20例正常人群样本，利用HLA-B*5801等位基因检测试剂盒进行检测，同时利用测序法对HLA-B等位基因高度突变的2/3/4外显子正反测序。灵敏度验证研究，随机选取1例阳性样本，按照HLA-B*5801等位基因检测试剂盒说明书要求稀释至最低浓度，检测结果与测序结果对比；抗干扰能力验证研究，随机选取3例阳性样本，加入规定要求的胆红素、脂质、阿司匹林作为干扰因素，利用PCR染料法检测，检测结果与测序结果对比。结果:45例痛风、高尿酸患者中有8例HLA-B*5801基因阳性，阳性率为17.8%;20例正常人群则有1例携带HLA-B*5801基因，阳性率5%。最低浓度样本也可正确检出结果，PCR染料法的准确性、灵敏性、特异性均为100%;3例阳性患者在胆红素、脂质等干扰物质影响下仍可正常检出，抗干扰能力良好。结论:PCR染料法可正确检测HLA-B*5801基因，灵敏度较好且在干扰物质存在的状况下结果不受影响，与测序结果一致，且操作更快捷方便。更多还原

【Abstract】 Objective: To evaluate the accuracy and sensitivity of fluorescent quantitative polymerase chain reaction(PCR) dye method for detecting the allele of human leukocyte antigen B locus 5801(HLA-B*5801) and analyze the results under interference factors, and to compare it with PCR-SBT sequencing method. Methods: 45 patients with gout, hyperuricemia and 20 normal people were collected and tested with HLA-B*5801 allele detection kit, while 2/3/4 exons with highly mutated HLA-B allele were sequenced by sequencing. In the sensitivity validation study, one positive sample was randomly selected and diluted to the lowestconcentration according to the instructions of HLA-B*5801 allele detection kit, and the test results were compared with the sequencing results; In the anti-interference verification study, three positive samples were randomly selected, bilirubin, lipid and aspirin were added as interference factors, PCR dye method was used for detection. The test results were aligned to the sequencing results. Results: HLA-B*5801 gene was positive in 8 of 45 patients with gout and hyperuricemia, the positive rate was 17.8%; One of the 20 normal people carried HLA-B*5801 gene, with a positive rate of 5%. The accuracy, sensitivity and specificity of PCR dye method are all 100%; Three positive patients could still be detected normally under the influence of bilirubin, lipid and other interfering substances, and their anti-interference ability was good. Conclusion: The PCR dye method can correctly detect the HLA-B*5801 gene, with good sensitivity and unaffected results in the presence of interfering substances, consistent with the sequencing results, and faster and convenient to operate.更多还原