PCR-SSP检测HLA-B27基因

HLA-B27 genotyping by polymerase chain reaction with sequence specific primers

目的 建立序列特异引物聚合酶链反应 (PCR SSP)检测HLA B2 7基因的方法。 方法 合成HLA B2 7基因和内对照人生长激素基因引物各一对 ,建立PCR SSP方法。提取 82例患者临床样品基因组DNA ,用于PCR扩增。其中 2 0例与微量淋巴细胞毒法 (MLCT)比较。 结果 PCR检测HLA B2 7阳性率为 6 8 3%。 2 0例样本同时行PCR SSP和MLCT法 ,PCR阳性 14例 (70 % ) ,MLCT法阳性 11例 (5 5 % ) ,两者一致率为 85 %。 结论 PCR SSP是一种敏感特异、快速简便的HLA B2 7检测方法 ,优于MLCT法。

Objective To develop HLA-B27 genotyping by polymerase chain reaction with sequence specific primers (PCR-SSP). Methods HLA-B27 genotyping of PCR-SSP was set up by synthesizing one pair specific primer of HLA-B27 gene and one pair intercontrol primer of human growth hormone gene. Genomic DNA of 82 patients were extracted and amplified. 20 samples were also tested by Microlymphocytotoxicity test (MLCT). Results 56 (68.3%) HLA-B27 positive samples were found by PCR-SSP. Of 20 samples,14 (75%) was the HLA-B27 positive by PCR-SSP and 11 (55%) by MLCT,and the coincident rate was 85%. Conclusion PCR-SSP is a sensitivity,specificity,rapid method,and is superior to the MLCT for genotype HLA-B27.