摘要
建立套式序列特异引物聚合酶链反应(PCR-SSP)方法,用于HLA-DR基因分型。利用一对外引物扩增HLA-DRB1第2外显子部分片段(283bp),作为第2次扩增的模板,PCR循环参数:98℃、5分钟;94℃、60秒,72℃、30秒,63℃、30秒,15个循环;利用各组特异性引物进行第2次扩增,产生与血清学相对应的HLA-DR型别,PCR循环参数:94℃、30秒,72℃、30秒,60℃、30秒,20个循环。结果表明本方法特异性好。在7个家庭20例受试者中进行了应用,与血清学方法比较,两方法确定出的型别,80%一致,20%不同,另有血清学为“空”的8个型别,基因分型全部为DR9阳性。
A nested PCR sequence specific primer (PCR-SSP) method was developed to be used in HLA-DR genotyping. A pair of primers was used to amplify a 283bp fragment of HLADRB1 exon 2, then 11 pairs of groupspecific primers were utilized to amplify the corresponding fragments by which HLA-DR specificities were determined. A total of 20 DNA samples from patients were analyzed for HLA matching before bone marrow transplantation by both serological and genetic methods. The results indicated that the same types were identified in 80%, different types in 20% by the two methods. In the 20 DNA samples, 7 kinds of types: DR2, DR36, DR4, DR7, DR9, DR11, DR12 were amplified.
出处
《中华器官移植杂志》
CAS
CSCD
1998年第3期137-139,共3页
Chinese Journal of Organ Transplantation