Goettner Gereon, Schulte-Spechtel Ulrike, Hillermann Ruth, Liegl Gabi, Wilske Bettina, Fingerle Volker
Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität München, Pettenkoferstr. 9a, D-80336 Munich, Germany.
J Clin Microbiol. 2005 Aug;43(8):3602-9. doi: 10.1128/JCM.43.8.3602-3609.2005.
We developed and evaluated a recombinant Borrelia line immunoblot assay based on 18 homologues of seven different antigens, i.e., p100, p58, p41i, BmpA, VlsE, OspC, and DbpA. Each recombinant antigen can be detected separately and is distinct even from homologues with identical molecular weights. This blot was compared to the recently described recombinant Borrelia Western immunoblot assay (U. Schulte-Spechtel, G. Lehnert, G. Liegl, V. Fingerle, C. Heimerl, B. J. Johnson, and B. Wilske, J. Clin. Microbiol. 41:1299-1303, 2003). To verify sensitivity and specificity, both blots were evaluated for reactivity with Borrelia-specific immunoglobulin G (IgG) and IgM antibodies with 85 sera from patients with different manifestations of Lyme borreliosis and 110 controls. According to European interpretation criteria for Borrelia Western blots, which define a serum as positive when it recognizes at least two bands, sensitivity increased significantly from 70.6% (Western blot) to 84.7% (line blot) for IgG (P = 0.042) and from 40.0% (Western blot) to 73.8% (line blot) for IgM (P < 0.005). The increased sensitivity for IgG detection is due to the new line blot technique, whereas the improvement in detection of IgM is mainly achieved through incorporation of the additional antigens. Notably, the recombinant VlsE of Borrelia garinii strain PBi displayed the highest sensitivity of all antigens tested for IgG detection and is also one of the most useful antigens for IgM. Due to its excellent sensitivity and specificity combined with ease of evaluation, this line immunoblot assay offers a useful improvement in serodiagnosis of Lyme borreliosis.
我们研发并评估了一种基于七种不同抗原(即p100、p58、p41i、BmpA、VlsE、OspC和DbpA)的18种同源物的重组疏螺旋体线性免疫印迹检测法。每种重组抗原都能被单独检测到,即使与分子量相同的同源物也有所不同。将这种印迹与最近描述的重组疏螺旋体免疫印迹检测法(U. Schulte-Spechtel、G. Lehnert、G. Liegl、V. Fingerle、C. Heimerl、B. J. Johnson和B. Wilske,《临床微生物学杂志》41:1299 - 1303,2003年)进行比较。为验证敏感性和特异性,用来自莱姆病不同临床表现患者的85份血清和110份对照血清,对两种印迹检测疏螺旋体特异性免疫球蛋白G(IgG)和IgM抗体的反应性进行评估。根据欧洲疏螺旋体免疫印迹的解读标准,当血清识别至少两条带时定义为阳性,IgG的敏感性从70.6%(免疫印迹法)显著提高到84.7%(线性印迹法)(P = 0.042),IgM的敏感性从40.0%(免疫印迹法)提高到73.8%(线性印迹法)(P < 0.005)。IgG检测敏感性的提高归因于新的线性印迹技术,而IgM检测的改善主要是通过加入额外的抗原实现的。值得注意的是,伽氏疏螺旋体菌株PBi的重组VlsE在所有检测的抗原中对IgG检测显示出最高的敏感性,也是IgM检测最有用的抗原之一。由于其出色的敏感性和特异性以及易于评估,这种线性免疫印迹检测法为莱姆病的血清学诊断提供了有益的改进。