HLA 基因序列电泳多态性分析及其在骨髓移植中的应用

PCR amplification of HLA genes followed by electrophoresis and its application in bone marrow transplantation

利用聚合酶链反应（ＰＣＲ）和聚丙烯酰胺凝胶电泳技术建立了一套人白细胞抗原（ＨＬＡ）配型技术，包括ＨＬＡ－ＤＲ单链构象多态性（ＰＣＲ－ＳＳＣＰ），异二聚体分析和ＨＬＡ－ＤＲ、－ＤＰＤＮＡ链构象多态性（ＰＣＲ－ＳＣＰ）分析技术。对６３例骨髓移植供、受体进行了分析，其中经血清学确定为ＨＬＡ相合的１５对，ＰＣＲ－ＳＣＰ分析完全相合；５对因病人血细胞数目和功能异常而供、受体血清学方法无法分型，ＰＣＲ－ＳＣＰ分析确定３对相合，２对不相合，病人临床缓解后重复血清学ＨＬＡ－Ａ，－Ｂ，－ＤＲ，－ＤＱ分型，证明ＳＣＰ分析结果正确；其余血清学不合的个体，ＰＣＲ－ＳＣＰ分析也不相合。６３例中有４２例进行了ＨＬＡ－ＤＲＰＣＲ－ＳＳＣＰ和异二聚体分析，结果与ＳＣＰ分析完全相同，其中９对相合，４对血清学结果无法判断。结果表明，三种技术都可准确地用于ＨＬＡ配型，但具有不同的特点。

In this study, three techniques, PCR SSCP, PCR SCP and heteroduplex analysis were developed and used in the selection of matched donor for BMT. A total of 63 patients and their potential donors were analyzed by PCR SCP. Of them, 42 were also tested with PCR SSCP and heteroduplex analysis. Of the 63 patients and their potential donors, 15 pairs of serologically matched patient donors and donor donors were also matched by HLA DR PCR SCP analysis. Five patient donor pairs with ambiguous serological typing results were three matched and two unmatched by HLA DR PCR SCP analysis. The results were verified later by serological examination once again in the period of clinical remission. The unmatched cases were also unmatched by PCR SCP. The typing results by PCR SCP were well identical to those by serological typing. Forty two of the 63 individuals were analyzed by HLA DR PCR SSCP and heteroduplex techniques and the result showed that 9 pairs of patients and their potential donors were matched and 4 pairs had ambiguous serological results. The results were identical to those by PCR SCP analysis. In comparison, all the three techniques can be used in HLA typing but with different methodological characteristics.