中国莱姆病伽氏疏螺旋体蛋白免疫印迹法诊断标准的研究

Standard criteria of Western blot for the diagnosis of Lyme disease caused by Borrelia garinii in China

目的研究中国莱姆病伽氏疏螺旋体基因型蛋白免疫印迹法(WB)的阳性诊断标准。方法采用中国莱姆病螺旋体伽氏疏螺旋体基因型参照菌株PD91作抗原,建立WB检测方法。采用Gel-Pro凝胶分析软件进行结果分析。病例组和对照组各组蛋白条带的频数分布比较采用χ2检验或Fisher′s精确检验,WB检测莱姆病阳性诊断标准确定采用ROC曲线。结果共检测127例莱姆病病例和504例对照的血清标本,经统计学分析得出我国莱姆病伽氏疏螺旋体检测中WB阳性诊断标准为:对于IgGP83/100、P58、P39、P30、OspC、P17、P66、OspA中至少有一条蛋白条带显色即可诊断为阳性,此标准敏感度为73.2%,特异度为99.4%;对于IgMP83/100、P58、OspA、P30、OspC、P17、P41中至少有一条蛋白条带显色则可诊断为阳性,此标准敏感度为50.4%,特异度为93.1%。结论建立了中国莱姆病伽氏疏螺旋体基因型WB阳性诊断标准,用于莱姆病的WB诊断具有较好的特异性。

Objective To study standard criteria of Western blot for the diagnosis of Lyme disease caused by Borrelia garinii in China. Methods To screen representative strain PD91 which attributes to predominant genospecies Borrelia garinii in Chinese Borrelia burgdoferi as the antigen, with SDS-PAGE, electrotransfer and immunoblot assays, and established a standard Western blot assay. The results were analyzed by Gel-Pro analysis software. The identity of groups were compared in 2 × 2 tables by X^2 analysis or Fisher＇s exact test. The criteria were estabhshed by ROC curve method. Results We tested 631 sera samples including 127 patients with Lyme disease and 504 controls. With Western blot for the diagnosis of Lyme disease in China, the following interpretation criteria were recommended： for IgG, at least one band of P83/100, P58, P39, P30, OspC, P17, P66 and OspA, the sensitivity was 73.2% and specificity was 99.4% ; for IgM, at least one band of P83/100, P58, OspA, P30, OspC, P17 and P41, the sensitivity was 50.4% and the specificity was 93.1%. Conclusion The standard criteria of Western blot for the diagnosis of Lyme disease showed a good specificity and sensitivity as proved our research.