PCR-SBT法和TaqMan探针法在HLA-E基因rs76971248和rs1264457位点分型中效果的比较及初步应用OA
Preliminary Application of PCR-SBT Method and TaqMan Probe Method in Genotyping of rs76971248 and rs1264457 Loci of HLA-E Gene
目的 对比PCR-SBT法与TaqMan探针法在HLA-E基因rs76971248(启动子区NT-26位)和rs1264457(第3外显子NT+756位)位点分型中的应用效果,分析两个SNP位点及对应基因型在健康献血者、血液肿瘤患者、实体瘤患者中的频率分布差异.方法 于深圳市血液中心收集208位健康献血者、111位血液肿瘤患者和100位实体瘤患者的全血,采用全自动核酸提取仪提取DNA,分别通过PCR-SBT法和TaqMan探针法进行HLA-E基因两个SNP位点分型,利用SPSS软件对不同人群的SNP位点及基因型频率进行卡方检验.结果 两种方法分型结果完全一致,但TaqMan探针法操作耗时更短,且无需产物纯化后测序步骤,更适用于大规模人群基因分型;两个SNP位点的频率在不同人群的结果显示,与健康献血者相比,血液肿瘤患者的rs76971248 T位点的频率要明显降低(P＜0.05,OR=0.39),而实体瘤患者无统计学差异(P＞0.05).而基因型结果显示,血液肿瘤患者rs76971248 G/G基因型和rs1264457 G/G基因型频率均高于健康献血者(P＜0.05,OR=2.43;P＜0.05,OR=1.98),而rs76971248 G/T基因型和rs1264457 A/G基因型频率低于健康献血者人群(P＜0.05,OR=0.40;P＜0.05,OR=0.54).实体瘤患者上述基因型频率与健康献血者无统计学差异(P＞0.05).结论 TaqMan探针法在HLA-E基因SNP快速分型中更具效率优势,可作为大规模肿瘤易感性筛查的优选技术.rs76971248 T位点是血液肿瘤的保护性位点,rs76971248 G/G基因型和rs1264457 G/G基因型是血液肿瘤的易感基因型.
Objective To compare the application effects of PCR-SBT and TaqMan probe method in genotyping of HLA-E gene rs76971248(promoter region NT-26)and rs1264457(exon 3 NT+756),and to analyze the differences in the frequency distribution of the two SNP loci and their corresponding genotypes among healthy blood donors,hematological tumor patients,and solid tumor patients.Methods Whole blood samples were collected from 208 healthy blood donors,111 hematological tumor patients and 100 solid tumor patients at Shenzhen Blood Center.DNA was extracted using a fully automatic nucleic acid extractor.Genotyping of the two SNP loci of HLA-E gene was performed by PCR-SBT and TaqMan probe method,respectively.Chi-square test was used to analyze the frequencies of SNP loci and genotypes among different populations using SPSS software.Results The genotyping results of the two methods were completely consistent,but the TaqMan probe method was less time-consuming to operate and did not require post-product purification and sequencing steps,making it more suitable for large-scale population genotyping.The frequency results of the two SNP loci among different populations showed that compared with healthy blood donors,the frequency of rs76971248 T in hematological tumor patients was significantly lower(P＜0.05,OR=0.39),while there was no statistical difference in solid tumor patients(P＞0.05).Regarding genotypes,the frequencies of rs76971248 G/G genotype and rs1264457 G/G genotype in hematological tumor patients were both significantly higher than those in healthy blood donors(P＜0.05,OR=2.43;P＜0.05,OR=1.98),whereas the frequencies of rs76971248 G/T genotype and rs1264457 A/G genotype were significantly lower than those in healthy blood donors(P＜0.05,OR=0.40;P＜0.05,OR=0.54).There were no statistical difference in the above genotype frequencies between solid tumor patients and healthy blood donors(P＞0.05).Conclusion The TaqMan probe method has greater efficiency advantage in the rapid genotyping of HLA-E gene SNP and can be used as a preferred technology for large-scale tumor susceptibility screening.The rs76971248 T allele is a protective locus for hematological tumors,and the rs76971248 G/G genotype and rs1264457 G/G genotype are susceptible genotypes for hematological tumors.
徐静;孙丽艳;洪文旭
广东药科大学公共卫生学院,广东 广州 510006深圳市血液中心,输血医学研究所,广东 深圳 518040广东药科大学公共卫生学院,广东 广州 510006||深圳市慢性病防治中心,广东 深圳 518020
医药卫生
HLA-E基因PCR-SBTTaqMan探针法血液肿瘤实体瘤
HLA-E genePCR-SBTTaqMan probe methodHematological tumorSolid tumor
《临床输血与检验》 2026 (2)
245-251,7
本课题受广东省医学科研基金项目(No.A2024098)、深圳市皮肤病防治研究所重点培育学科(No.SZDKCD001)资助
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