PCR-SSP法HLA-A、B基因分型与血清学分型的比较

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (11): 1267-1270.

PCR-SSP法HLA-A、B基因分型与血清学分型的比较

武大林¹, 凌汉新², 唐浩¹

1. 南方医科大学南方医院医学中心实验科, 广东, 广州, 510515;
2. 广州市第十二人民医院肾内科, 广东, 广州, 510620

出版日期:2004-11-20 发布日期:2004-11-20
基金资助:
收稿日期:2003-5-26。
基金项目:第一军医大学南方医院1998新技术攻关项目(98字第011号)
作者简介:武大林(1954- ),女,1986年毕业于第三军医大学,主任检验技师,电话:020-85141042,E-mail:wksys@fimmu.com>

Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers

WU Da-lin¹, LING Han-xin², TANG Hao¹

1. 南方医科大学南方医院医学中心实验科, 广东, 广州, 510515;
2. 广州市第十二人民医院肾内科, 广东, 广州, 510620

Online:2004-11-20 Published:2004-11-20

摘要/Abstract

摘要： 目的比较聚合酶链反应-序列特异性引物(PCR-SSP)进行HLA-Ⅰ类A、B抗原位点分型的准确性,并探讨血清学分型错误发生的原因。方法用PCR-SSP以及单克隆抗体血清学分型技术对HLA-A、B分型并比较。结果 34例样本PCR-SSP基因分型无假阳性和假阴性出现。PCR-SSP法与血清学比较,血清学检出错误或漏检率分别为HLA-A位点23.5%,B位点26.5%。血清学发生错误或易混淆的抗原有:A2和A68、A32和A33,B5、B60和61。结论 PCR-SSP法进行HLA-A、B抗原等位基因分型具有分辨率高、特异性强、重复性好、实验过程简捷快速、分型结果较血清学更加准确可靠的优点。

Abstract: 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.Conclusion s DNA typing for HLA-Ⅰclass (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.

中图分类号:

R446.11

武大林¹, 凌汉新², 唐浩¹. PCR-SSP法HLA-A、B基因分型与血清学分型的比较[J]. 南方医科大学学报, 2004, 24(11): 1267-1270.

WU Da-lin¹, LING Han-xin², TANG Hao¹. Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers[J]. Journal of Southern Medical University, 2004, 24(11): 1267-1270.

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