采用 PCR-SSP法检测HLA-B27基因

目的：建立顺序特异引物聚合酶链反应（ＰＣＲ－ＳＳＰ）方法检测ＨＩＡ－Ｂ２７基因并与血清学方法比较。方法：设计合成Ｂ２７特异物３个和内源性阳性对照２个，建立ＰＣＲ－ＳＳＰ方法，用于Ｂ２７基因分型。血清学分型为一步法单克隆抗估方法。临床样本１００份，不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：１００例临床样本ＰＣＲ－ＳＳＰ行Ｂ２７基因分型均获成功。总耗时４ｈ。其中Ｂ２７阴性５８例，阳

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

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.