PCR-SSP及PCR-SSCP在骨髓移植配型中的联合应用

Combined application of PCRSSP and PCRSSCP in bone marrow transplantation typing

目的研究ＨＬＡ抗原分型的方法。方法对２０对已进行ＨＬＡ抗原血清学分型的骨髓移植供、受者采用序列特异引物聚合酶链反应（ＰＣＲＳＳＰ）及单链构象多态性聚合酶链反应（ＰＣＲＳＳＣＰ）进行ＨＬＡ抗原的基因分型。ＤＮＡ的提取采用快速盐析法。结果基因分型结果与血清学分型一致者有１８对，基因分型能明确判定而血清学分型无法判断者有２对；ＰＣＲＳＳＰ与ＰＣＲＳＳＣＰ结果完全一致者有１９对；１对ＰＣＲＳＳＰ分型为ＨＬＡＤＲＢ１相合而ＤＱＢ１不相合，ＰＣＲＳＳＣＰ分型为ＨＬＡＤＲＢ１有轻度差别，而ＤＱＢ１、ＤＰＢ１有明显差别，该例患者骨髓移植后发生了Ⅳ级移植物抗宿主病。结论ＰＣＲＳＳＰ与ＰＣＲＳＳＣＰ联合应用有利于ＨＬＡ配型的准确性。

Objective To study the method for human leukocyte antigen (HLA) typing. Methods HLA genotyping was performed by using PCRSSP and PCRSSCP in 20 pairs of bone marrow transplantation donorrecipients who underwent serologic typing. DNA from 20 pairs of samples were extracted by a fast salting out method. Results The serological and genotyping results were identical in 18 cases, serological typing was failure in 2 cases who could be typed easily by genotyping methods. Between the two genotyping methods, there were 19 cases of identical results and 1 case of different results. After the patient received the graft, he got grade graft versus host disease (GVHD). Conclusion The combined application of PCRSSP and PCRSSCP was an ideal typing method and might be helpful to accurately tpye HLA.