中国输血杂志2026，Vol.39Issue(5)：596-602,7.DOI:10.13303/j.cjbt.issn.1004-549x.2026.05.004
采用KASP技术进行RhD阳性患者RHCE基因分型的临床应用研究
Clinical application of KASP-based RHCE genotyping in RhD-positive patients
摘要
Abstract
Objective To develop a RHCE genotyping assay based on kompetitive allele-specific PCR(KASP)and as-sess its clinical accuracy for RhCE blood group determination.Methods KASP primers were designed to interrogate three RHCE loci:the 109 bp insertion/deletion in intron 2,c.307T>C,and c.676C>G.A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology.Discordant samples(n=10)were retested by both methods and further resolved by Sanger sequencing.An additional 377 cases were tested for the c.48C>G locus to eval-uate the predictive accuracy of individual loci and combined locus testing for RhC antigen.Results Genotyping concord-ance with serology was 100.0%for both the c.676C>G locus(RhE/Rhe)and the c.307T>C locus(Rhc).For RhC predic-tion using the 109 bp insertion,overall accuracy was 99.7%(1 191/1 194);the 3 discordant cases were confirmed by San-ger sequencing to be false negatives attributable to 109 bp deletion in intron 2.Testing the c.48C>G allele for RhC predic-tion yielded 7 false positives,with an accuracy of 98.1%(370/377).RhC antigen status was determined by combining the 109 bp insertion and the c.48C allele.After excluding 10 samples with inconsistent results between the two loci,the accu-racy reached 100%in the remaining 367 samples.When both loci were applied in combination,accuracy reached 100%in the 367 cases with concordant results.Among the 1 194 patients,CCee(45.8%)and CcEe(31.7%)were the most com-mon RhCE phenotypes.The e antigen had the highest positivity rate(92.2%),and the Ce haplotype was the most frequent(66.9%).Conclusion The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing.Combining the 109 bp insertion/deletion with the c.48C allele significantly improves RhC antigen prediction compared with either locus alone.This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu,providing lo-cal RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.关键词
RHCE/基因分型/竞争性等位基因特异性聚合酶链式反应/血型抗原Key words
RHCE/genotyping/kompetitive allele-specific PCR/blood group antigens分类
医药卫生引用本文复制引用
连晓玉,徐海霞,王珏,李玲,刘忠,李梦丹,官笑羽,田力,王陈赢,吴迪,罗天琼,杜晓林,纪欣..采用KASP技术进行RhD阳性患者RHCE基因分型的临床应用研究[J].中国输血杂志,2026,39(5):596-602,7.基金项目
成都市科技局重点研发支撑计划成果转化示范项目(2024-YF09-00052-SN) （2024-YF09-00052-SN）
中国医学科学院医学与健康科技创新工程(2021-I2M-1-060) （2021-I2M-1-060）
