PCR-SSP检测HLA-B27基因的方法及临床意义

HLA-B27 typing by PCR-SSP and by microlymphocytoxicity test

目的 :建立快捷、特异的PCR SSP检测HLA B2 7基因的方法。方法 :以钾缓冲液提取 5 0～ 2 0 0 μl外周血中的DNA ,设计序列特异性的HLA B2 7引物进行PCR扩增检测HLA B2 7基因 ,结果与淋巴细胞毒方法 (MLCT)相对比。结果 :所设计引物可扩增出目的基因片段 ,钾缓冲液提取DNA可满足PCR要求 ,全部检测过程可在 6h内完成。在 2 2 5例门诊和住院患者中 ,HLA B2 7阳性率在PCR方法和MLCT方法分别为 77 3%和 6 8 0 % ,两者的一致率为 83 6 %。在 88例肯定的脊柱关节病 (SpAs)住院患者中 ,HLA B2 7阳性率在PCR方法为 92 1% ,在MLCT方法为 81 8% ,两者的一致率为 89 8%。结论 :PCR方法检测HLA B2 7较MLCT方法敏感、特异和快捷 ,标本用量少、采集和保存方便 。

Objective:To develop HLA B27 typing by a group specific polymerase chain reaction(PCR) amplification with a sequence specific primer pair(PCR SSP).Methods:A new and simple DNA extraction method from 200μl peripheral blood sample was used and a HLA B27 sequence specific primer pair for exon 2 of the HLA B27 gene based HLA class I antigen sequence were designed Two hundred and twenty five DNA samples from patients with spondyloarthropathies(SpAs) and other arthritis were amplified by PCR with B27 SSP All samples were also tested by Microlymphocytotoxicity test(MLCT).Results:174 (77 3%) HLA B27 positive samples were found by PCR, 153 (68 8%) by MLCT with a coincident rate of 83 9% In 88 inpatients with definite SpAs, the HLA B27 positive rate was 92 1% by PCR and 81 8% by MLCT, and the coincident rate was 89 8% The positive rate of PCR method was 10 3% higher than that of MLCT DNA extraction and PCR amplifying could be finished within 6 hours. Conclusion:PCR is superior to the MLCT to determine HLA B27 The method we described is sensitive, specific, simple and rapid, particularly when large numbers of samples are being studied.