HLA—Ⅱ血清学分型与微量SSP法基因分型的比较分析

目的 验证微量序列特异性引物（SSP）技术进行HLA－Ⅱ类移植配型的准确性并探讨血清学分型错误发生的原因。方法 采用聚合酶链反应结合微量SSP技术（简称微量PCR－SSP法）以及单克隆抗体血清学分型技术对血清样品进行HLA分型并比较其结果。结果 （1）微量PCR－SSP法检测的110例样本中，99例检出HLA－DR的396个等位基因、11例检出HLA－DQ的22个等位基因，检出10％的单倍型个体；（2）两种方法对比研究发现单克隆抗体血清学分型检出错误和漏检率较高，其误差在DR和DQ分别为38．81％和50．75％；（3）错误发生和易混淆的抗原为：DR15／16、11／12、13／14、8、12和DQ5／6、8／9。结论 微量SSP法可以准确检定血清学易漏检和发生错误的抗原等位基因。

Comparative analysis of serologic typing and HLA-II typing by micro-PCR-SSP.]

OBJECTIVE: To evaluate the accuracy of polymerase chain reaction with sequence specific primers (PCR-SSP) in HLA-II genotyping and analyze the causes of the errors occurring during the genotyping. METHOD: Blood samples were obtained form patients with chronic renal insufficiency, leukemia or thalassemia and also from normal subjects. HLA-DR and -DQ genotyping of the sera from the 110 subjects was performed using micro-PCR-SSP and comparison was made with the results obtained from monoclonal antibody serologic typing. RESULT: Of the 110 samples detected by micro-PCR-SSP, 396 alleles of HLA-DR were identified in 99 cases and 22 of HLA-DQ in 11 cases, and 10% of the subjects were identified as homozygote individuals. Examination by both of the 2 methods in 67 cases indicated high rates of missed diagnoses and misdiagnoses by serologic typing with the diagnostic discrepancy as high as 38.81% and 50.75% for HLA-DR and -DQ respectively. The antigens DR 15/16, 11/12, 13/14, 8 or 12; DQ 5/6, 8/9 were among those that frequently gave rise to errors or confusion. CONCLUSION: Micro-PCR-SSP method can accurately detect the alleles of HLA-II antigens that are easy to be missed or mistaken by serological typing method.