[血清学分型与应用序列特异性引物PCR技术进行HLA - A、B抗原基因分型的比较研究]
[Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers].
作者信息
Wu Da-lin, Ling Han-xin, Tang Hao
机构信息
Central Laboratory, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
出版信息
Di Yi Jun Yi Da Xue Xue Bao. 2004 Nov;24(11):1267-70.
OBJECTIVE
To evaluate the accuracy of PCR with sequence-specific primers (PCR-SSP) for HLA-I genotyping and analyze the causes of the errors occurring in the genotyping.
METHODS
DNA samples and were obtained from 34 clinical patients, and serological typing with monoclonal antibody (mAb) and HLA-A and, B antigen genotyping with PCR-SSP were performed.
RESULTS
HLA-A and, B alleles were successfully typed in 34 clinical samples by mAb and PCR-SSP. No false positive or false negative results were found, and the erroneous and missed diagnosis rates were obviously higher in serological detection, being 23.5% for HLA-A and 26.5% for HLA-B. Error or confusion was more likely to occur in the antigens of A2 and A68, A32 and A33, B5, B60 and B61.
CONCLUSIONS
DNA typing for HLA-I class (A, B antigens) by PCR-SSP has high resolution, high specificity, and good reproducibility, which is more suitable for clinical application than serological typing. PCR-SSP may accurately detect the alleles that are easily missed or mistaken in serological typing.
目的
评估序列特异性引物聚合酶链反应（PCR-SSP）对HLA-I基因分型的准确性，并分析基因分型中出现错误的原因。
方法
从34例临床患者中获取DNA样本，采用单克隆抗体（mAb）进行血清学分型，采用PCR-SSP进行HLA-A和B抗原基因分型。
结果
通过mAb和PCR-SSP成功对34例临床样本进行了HLA-A和B等位基因分型。未发现假阳性或假阴性结果，血清学检测中的错误诊断率和漏诊率明显更高，HLA-A为23.5%，HLA-B为26.5%。在A2和A68、A32和A33、B5、B60和B61抗原中更容易出现错误或混淆。
结论
PCR-SSP对HLA-I类（A、B抗原）进行DNA分型具有高分辨率、高特异性和良好的重复性，比血清学分型更适合临床应用。PCR-SSP可准确检测血清学分型中容易漏检或误判的等位基因。
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