Wu Da-Lin, Ling Han-Xin, Ding Hong, Zhang Yi
Department of Transfusion, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2002 Mar;22(3):247-9.
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.
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.
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.
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.
评估序列特异性引物聚合酶链反应（PCR-SSP）在HLA-II基因分型中的准确性，并分析基因分型过程中出现错误的原因。
采集慢性肾功能不全、白血病或地中海贫血患者以及正常受试者的血样。采用微PCR-SSP对110名受试者血清进行HLA-DR和-DQ基因分型，并与单克隆抗体血清学分型结果进行比较。
微PCR-SSP检测的110份样本中，99例鉴定出396个HLA-DR等位基因，11例鉴定出22个HLA-DQ等位基因，10%的受试者被鉴定为纯合子个体。67例样本采用两种方法检测，结果显示血清学分型漏诊和误诊率较高，HLA-DR和-DQ的诊断差异分别高达38.81%和50.75%。DR 15/16、11/12、13/14、8或12；DQ 5/6、8/9等抗原是经常导致错误或混淆的抗原。
微PCR-SSP方法能够准确检测出血清学分型容易漏检或误诊的HLA-II抗原等位基因。
