采用PCRSSP法检测HLA-B27基因

Genotyping for HLA-B27 by polymerase chain reaction with sequence specific primers

目的 :建立顺序特异引物聚合酶链反应 (PCR SSP)方法检测HLA B2 7基因并与血清学方法比较。方法 :设计合成B2 7特异引物 3个和内源性阳性对照 2个 ,建立PCR SSP方法 ,用于B2 7基因分型。血清学分型为一步法单克隆抗体方法。临床样本 10 0份 ,来源于可疑强直性脊柱炎患者。快速酚氯仿法提取模板DNA。结果 :10 0例临床样本PCR SSP行B2 7基因分型均获成功。总耗时 4h。其中B2 7阴性 5 8例 ,阳性 42例 ,5例临床诊断为强直性脊柱炎 ,占 11 9%。 30份样本同时行血清学分型 ,2 0份B2 7阴性 ,5份阳性 ,5份可疑阳性。其中 2 0份阴性中 3例为阳性 ,5份可疑阳性中 2例为阴性。结论 :PCR SSP行B2 7基因分型 ,分辨度高、特异性强 ,快捷、准确 ,适合于临床应用。

Objective: Genotyping for HLA B27 by polymerase(PCR SSP)was established and compared with serology.Methods:Genotyping for B27 of PCR SSP method was set up by designing and synthesizing 3 specific priers and 2 control priers according to the HLA B27 nucleotide sequences.Serology was a one step monoclonal antibody typing procedure.A total of 100 samples from ambiguous ankylosing spondylitis were entered into the study.Genomic DNA was obtained from rapid phenolchloro form extraction. Results:Genotyping for B27 was successfully typed in 100 samples by PCR SSP,containing 58 with B27 negative,42 with B27 positive.5 patients with B27 positive(11.9%) were diagnosed ankylosing spondylitis.30 samples was typed by serology,including 20 with B27 negative 5 positive and 5 ambiguous positive.3 out of 20 negative were positive and 2 out of 5 ambiguous positive were negative by PCR SSP typing for HLA B27.Conclusion:Genotyping for HLA B27 by PCR SSP has proved to be a high sensitivity,high specificity,rapid and accurate technique,suitable for clinical application.