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摘要: 目的 探讨实时荧光PCR方法用于中国人群RHc基因检测的可行性。方法 收集144例大连地区RhD阳性健康献血者血样，使用血清学分型方法检测样本RhCcEe血型表型。使用检测RHCE基因第2外显子178C>A和307C>T多态性位点的TaqMan探针实时荧光PCR方法对血液标本进行RHc基因分型，并将基因分型结果与血清学分型结果相比较。结果 经血清学检测，144例样本中有60例为CCee、48例为CcEe、17例为ccEE、11例为Ccee、7例为ccEe、1例为ccee；经TaqMan探针实时荧光PCR方法检测，发现RHc基因阳性为84例、阴性为60例，基因分型结果与血清学分型结果一致，该方法的灵敏度、特异度和准确度均为100%。结论 TaqMan探针实时荧光PCR方法适用于检测中国人群RHc基因，可用于临床实验室对Rhc血型进行基因检测。
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关键词:
- RHc基因 /
- TaqMan探针实时荧光PCR /
- 基因分型
Abstract: Objective To determine the feasibility of RHc genotyping through real-time PCR in Chinese population.Methods A total of 144 participants were recruited from RhD positive healthy blood donors in Dalian. RhCcEe phenotypes of the samples were tested by serological test. RHc of the samples were genotyped by targeting exon 2 178C>A and 307C>T mutation of the RHCE gene through real-time PCR with TaqMan probe. The results of genotyping and serological typing were compared.Results Serological test showed that 60 cases were CCee, 48 cases were CcEe, 17 cases were ccEE, 11 cases were Ccee, 7 cases were ccEe and 1 case was ccee among 144 samples. By genotyping through real-time PCR with TaqMan probe, RHc gene was positive in 84 cases and negative in 60 cases. The results of genotyping and serological typing were consistent. The sensitivity, specificity and accuracy of the PCR method were 100%.Conclusion Real-time PCR with TaqMan probe might be feasible in RHc genotyping in Chinese population, which could be used for gene detection of Rhc blood group in clinical laboratory.-
Key words:
- RHc gene /
- real-time PCR with TaqMan probe /
- genotyping
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表 1 TaqMan探针实时荧光PCR探针与引物序列
探针/引物名称 序列(5′-3′) RHc_F 5′-TGGGCTTCCTCACCTCAAA-3′ RHc_R ′5-TGATGACCACCTTCCCAGG-3′ RHc_P 5′-(FAM)CAATCCTGCTGGACGGCTTCCTGA(BHQ1)-3′ GAPDH_F 5′-CCCCACACACATGCACTTACC-3′ GAPDH_R 5′-CCTAGTCCCAGGGCTTTGATT-3′ GAPDH_P 5′-(TAMRA)AAAGAGCTAGGAAGGACAGGCAACTTGGC(BHQ2)-3′ 
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表 2 RHc基因分型结果与血清学分型结果比较
(n=144) 血清学分型 例数 RHc基因分型结果 CCee 60 - Ccee 11 + CcEe 48 + ccee 1 + ccEE 17 + ccEe 7 +
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