应用PCR-SSP方法检测HLA-B27亚型及其临床意义

Clinical implication of HLA-B27 subtypes by polymerase chain reaction with sequence specific primers

[目的]通过PCR-SSP方法对HLA-B27亚型进行检测,探讨其与强直性脊柱炎(AS)的相关性,并对大连地区B-27分型情况进行研究。[方法]对40例确定为HLA-B27阳性的样本(临床诊断28例AS,12例非AS)采用序列特异性引物-聚合酶链反应(PCR-SSP)检测方法进行HLA-B27亚型分型。[结果]40例样本用PCR-SSP方法检测均获成功。共检出4种亚型:B2704、B2705、B2702和B2710。B2704 23例占57.5%(23/40例);B2705 14例占35%(14/40例);B2702 2例占5%(2/40例);B2710 1例占2.5%(1/40例)。28例AS标本与12例非AS标本中B2704、B2705所占比例统计学分析无显著差异(B2704χ2=0.5894,P>0.05;B2705χ2=0.02,P>0.05)。[结论]PCR-SSP方法对HLA-B27基因分型,分辨度高、特异性强、快捷、准确,适合于临床应用。HLA-B27亚型的检测可作为强直性脊柱炎诊断(AS)的重要的参考指标。大连地区HLA-B27亚型以B2704和B2705为主并出现1例B2710型。

[Objective] To detect the HLA - B27 subtypes by Polymerase Chain Reaction with sequence specific primers. To study the clinical meaning of HLA - B27 subtypes associated with ankylosing spondylitis （AS） and investigate HLA - B27 subtypes of the Dalian area. [Methods] Subtyping for HLA - B27 of Polymerase Chain Reaction with sequence specific primers（PCR - SSP） method was applied. A total of 40 HLA - B27 positive samples were entered into the study, containing 28 samples diagnosed ankylosing spondylitis and 12 samples diagnosed not ankylosing spondylitis. [Results] Subtyping for B27 was successfully tested in 40 samples by PCR - SSP. Checkout four HLA - B27 Subtypes： B2704, B2705, B2702 and B2710. B2704 accounting for 57.5% in all 40 samples, B2705 accounting for 35%. There was no significant difference between the rate of B2704, B2705 positive sample in ankylosing spondylitis and not ankylosing spondylitis （B2704χ^2 = 0.5894, P 〉 0.05; B2705χ^2 = 0.02, P 〉 0.05）. [Conclusion] Subtyping for HLA - B27 by PCR - SSP has proved to be a high - sensitivity, high - specificity, rapid and accurate technique, suitable for clinical application. It is necessary to diagnose ankylosing spondylitis in clinic. The dominating subtype for HLA - B27 of Dalian area is B704 and B2705, only finding B2710 one example.