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目的采用顺序特异引物聚合酶链反应技术（ＰＣＲＳＳＰ）建立汉族人群白细胞抗原Ⅰ（ＨＬＡⅠ）类抗原Ａ、Ｂ位点ＤＮＡ分型方法。方法设计合成８１个特异性引物和１对阳性对照引物，组成２０个ＰＣＲ反应用于Ａ位点分型、４１个ＰＣＲ反应用于Ｂ位点分型，建立一步法ＰＣＲＳＳＰ方法，应用于６２份标准ＤＮＡ和３４５例肾移植供受者的ＨＬＡⅠ类ＤＮＡ分型。结果所有样本ＰＣＲＳＳＰ基因分型均获得成功，无假阳性和假阴性出现，４０份样本的重复率１００％，总耗时５小时。分型结果经双盲验证完全符合。可准确分辨出Ａ位点等位基因１９个、Ｂ位点等位基因４１个；实际检出汉族人群Ａ抗原特异性１３个、Ｂ抗原特异性３２个。结论ＰＣＲＳＳＰ技术行ＨＬＡⅠ类Ａ、Ｂ位点ＤＮＡ分型，分辨率高、特异性强、重复性好、相对简捷快速，分型结果较血清学方法更加精确可靠，适合于临床应用
Objective To establish a DNA typing method for leukocyte antigenⅠ (HLAⅠ) antigen A and B sites in Han population by PCRSSP with sequence-specific primers. Methods 81 specific primers and 1 pair of positive control primers were designed and synthesized. Twenty PCR reactions were designed for A site typing and 41 PCR reactions were used for site B typing. One-step PCR-SP method was established and applied to 62 standard DNA and 345 cases of renal transplantation for HLA class Ⅰ DNA typing. Results PCR-SSP genotyping was successful in all samples without any false-positive or false-negative results. The repeatability of the 40 samples was 100% and the total time was 5 hours. The typing results were fully validated by double-blind validation. 19 alleles at A locus and 41 alleles at loci B can be distinguished accurately. Thirteen A antigen specificities and 32 B antigen specificities were detected in Han population. Conclusion PCR SP technique HLA class Ⅰ A, B site DNA typing, high resolution, specificity, good reproducibility, relatively simple and rapid, the typing results than serological methods more accurate and reliable, suitable for clinical application