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目的 探讨血清学分型方法在脐血HLA分型中的应用价值。方法 对 180 0份脐血的HLA AB分型结果进行分析 ,并与其中随机抽取的 36份标本的聚合酶链反应 序列特异性引物 (PCR SSP)分型结果进行比较分析。血清学分型分别以T免疫磁珠和淋巴细胞分层液分离细胞。结果 T免疫磁珠分离的T淋巴细胞比淋巴细胞分层液分离的单个核细胞活性更好 ;体外搁置时间较长 (2 4～48h)的脐血须用T免疫磁珠分离细胞 ;与外周血单个核细胞相比 ,脐血细胞的血清学反应强度偏低。除 2 8份因细胞分离后量少或活性低外 ,1772份脐血标本的HLA AB血清学分型中 17.5 % (310份 )有不能确定的抗原 ,其主要原因依次为交叉反应及部分分型试剂特异性不高、部分抗原反应弱、细胞活性低所致的高本底及非特异反应。 36份标本PCR SSP分型均获成功 ,与之对照 ,血清学分型有 16 .8%的错误率 ,13.9%分辨率低于PCR SSP分型。结论 血清学分型方法虽快速简便 ,但有一定的误差 ,用基因分型作为补充是必要的。
Objective To investigate the value of serological typing in HLA typing of umbilical cord blood. Methods The HLA-AB typing results of 180 0 cord blood samples were analyzed and compared with PCR SSP typing results of 36 randomly selected samples. Serotypes were separated by T immunomagnetic beads and lymphocyte stratification. Results T lymphocytes isolated from T magnetic beads were more active than mononuclear cells isolated from lymphocyte stratified fluid. Umbilical cord blood with long shelf-life in vitro (24h ~ 48h) Serum mononuclear cells compared to umbilical cord blood cells seroconversion intensity is low. Except for 28 samples, 17.5% (310 copies) of HLA-AB serological classification of 1772 cord blood samples had undetermined antigens due to the small amount or low activity after cell separation. The main reasons were followed by cross-reaction and partial classification Reagent specificity is not high, part of the antigen response is weak, low cell activity caused by high background and non-specific reaction. In all 36 samples, PCR SSP typing was successful. In contrast, the serological typing rate was 16.8%, and the 13.9% resolution was lower than PCR SSP typing. Conclusion Although the serological typing method is quick and easy, it has some errors. It is necessary to use genotyping as a supplement.