摘要
目的 建立一种人白细胞抗原 (HLA)基因分型方法。方法 用序列特异性引物和荧光染料SYBRGreenⅠ聚合酶链式反应 (PCR)方法 ,对 1份已知HLA DRB1 12的标本和 12份未知型别的临床标本进行HLA DRB1 12位点检测和分型。结果 12份标本的扩增产物经熔解曲线分析 ,有 3份标本为DRB1 12 ,产物用离心柱纯化后 ,直接进行DNA测序 ,结果证实为DRB1 12阳性。结论 SYBRGreenⅠPCR操作简便 ,结果准确 。
Objective To detect the HLA DRB1*12 by use of a new genotyping method for HLA SYBR Green Ⅰ PCR. Methods 1 HLA DRB1*12 sample and 12 unknown clinical sample were collected. The typing of these samples were determined by PCR, which includes a fluorescence dye, SYBR GreenⅠ,and the sequence specific primer. SYBR GreenⅠcould bind to the dsDNA to exhibit fluorescence. The fluorescence enhancement depends on the accumulation of the amplified product. Results 3 of the 12 unknown sample was proved to be DRB1*12 by the analysis of the melting curve of the amplified products. After purification, the type of these products were confirmed DRB1*12 by DNA based sequencing. Conclusions The results of SYBR GreenⅠPCR demonstrate that it can be a new strategy for HLA typing because of its convenience and accuracy.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2003年第10期597-599,共3页
Chinese Journal of Laboratory Medicine
