脐血HLA-Ⅰ类抗原血清学分型与基因分型的比较研究.docVIP

???? 脐血HLA-Ⅰ类抗原血清学分型与?基因分型的比较研究 ???? ????　【摘要】　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1800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(24～48h)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28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。　　【关键词】　胎血；　HLA抗原；　血清学分型；　基因分型 Comparison of serological and DNA typings for HLA-AB of cord blood samples LI Dan，LIU Haiyan，YU Yan， et al.（Peoples Hospital, Beijing Medical University，Beijing　100044，China） 　　【Abstract】　Objective　To evaluate the practical value of serological method for HLA-AB typing of cord blood samples. Methods　Serological typing results of 1800 cord blood samples were analyzed and 36 of them were compared with that of PCR-SSP (polymerase chain reaction-sequence specific primers). Lymphocytes in cord blood samples were isolated by T-immunomegnetic beads or Ficoll. Results　T lymphocytes isolated by fluorobeads have better activity than that in mononuclear cells isolated by Ficoll. In order to get the more active cord blood cells, samples stored in vitro over 24 hours must be isolated with fluorobeads. Its serological reaction is lower as compared with that of peripheral mononuclear cells. Except 28 samples which had too many dead cells or few cells, there were 310(17.5%) unconfirmed antigens among 1772 cases by serological typing. The main causes were low specificity of typing reagents, cross reaction, weak serological reaction, high background of dead cells and extra reaction. Confirmed by PCR-SSP, the serological typing had a wrong result rate of 13.88%. Conclusion　Although serological typing is rapid and easy, it is necessary to use genetic typing as a complement to it.　　【Key words】　Cord blood；　HLA，human；　Serologic typing；　Genetic typing 　　脐血含有丰富的造血干细胞、来源广泛,且采集对供体不会造成伤