应用Taq Man探针法与高分辨熔解曲线法检测HLA-B27基因诊断强直性脊柱炎的价值

目的运用Taq Man探针法与高分辨熔解曲线法检测疑似强直性脊柱炎患者的人类白细胞抗原HLA-B27,并探讨其方法的有效性和应用价值。方法分别将疑似强直性脊柱炎患者的血样DNA进行HLA-B27的Taq Man探针法和高分辨熔解曲线法检测,并以DNA直接测序法所得阳性结果作为金标准,来确定Taq Man探针法和高分辨熔解曲线法的敏感度、特异度、阳性预测值和阴性预测值。结果 55例样本中,Taq Man探针法和高分辨熔解曲线法与DNA测序法的符合率分别为98.2%和92.7%。Taq Man探针法检测HLA-B27的敏感度为0.970,特异度为1.000,阳性预测值为1.000,阴性预测值为0.956;高分辨熔解曲线法检测HLA-B27的敏感度为0.879,特异度为1.000,阳性预测值为1.000,阴性预测值为0.846。结论 Taq Man探针法与高分辨熔解曲线法均具有准确性好、灵敏度高、检测快速等优点,适合于临床HLA-B27的检测。

Value of application of Taq Man PCR and high-resolution melting curve method in detection of HLA-B27 for ankylosing spondylitis diagnosis

[Objective]To validate the method of Taq Man PCR and high resolution melting curve analysis(HRM) for rapid detecting HLA-B27 in suspected ankylosing spondylitis(AS) patients,and investigate its feasibility and application value. [Methods]DNA samples of AS patients were examined by Taq Man PCR and HRM,respectively.The positive result of DNA sequencing were taken as gold standard for the confirmation of the sensitivity,specificity,positive predictive value and negative predictive value of this method. [Results]Of 55 DNA samples,the coincidence rates were 98.2% and 92.7% obtained by Taq Man PCR and HRM to DNA sequencing.The sensitivities,specificities,positive predictive value and negative predictive value of Taq Man PCR and HRM detecting HLA-B27 were 0.970,1.000,1.000,0.956;0.879,1.000,1.000,0.846,respectively. [Conclusion]Taq Man PCR and HRM providing accurate,specific and rapid detection of HLA-B27,it can be applied for the detection of HLA-B27 in clinic.