Wilske B, Habermann C, Fingerle V, Hillenbrand B, Jauris-Heipke S, Lehnert G, Pradel I, Rössler D, Schulte-Spechtel U
Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität München, Munich, Germany.
Med Microbiol Immunol. 1999 Dec;188(3):139-44. doi: 10.1007/s004300050116.
We have previously described the use of the following recombinant antigens for serodiagnostic immunoblots: p83/100, p39, OspC and p41 (flagellin) internal fragment [Wilske et al. (1993) Med Microbiol Immunol 182:255-270; Rossler et al. (1997) J Clin Microbiol 35:2752-2758]. In our currently used immunoblot p83/100 is derived from strain PKo (Borrelia afzelii), p39 (BmpA) and OspC from strains PKa2 (B. hurgdorferi sensu stricto), PKo and PBi (B. garinii), respectively; the p41 (flagellin) internal fragments were cloned from strains PKo and PBi. In this study we describe the use of two additional recombinantly expressed highly immunogenic proteins Osp 7 (derived from PKo) and p58 (derived from PBi). A clinically well-defined panel of sera from 147 Lyme borreliosis patients and 139 controls previously tested by a standardized whole cell lysate immunoblot [Hauser et al. (1997) J Clin Microbiol 35:1433-1444] was investigated in the recombinant immunoblot without (old recombinant immunoblot) and with Ospl7 and p58 (new recombinant immunoblot) for IgG antibodies. The sensitivity of the recombinant IgG immunoblot for diagnosis of stage II and stage III could be significantly improved by addition of Osp17 and p58 without loss of specificity. With the exception of sera from patients with erythema migrans the diagnostic sensitivity is comparable to the whole cell lysate IgG immunoblot. The main advantage of the recombinant immunoblot is the easy identification of diagnostic bands, whereas the identification of bands in the whole cell lysate immunoblot is difficult. The recombinant immunoblot is especially suitable where large series of sera need to be investigated.
p83/100、p39、OspC和p41（鞭毛蛋白）内部片段[Wilske等人（1993年）《医学微生物学与免疫学》182:255 - 270；Rossler等人（1997年）《临床微生物学杂志》35:2752 - 2758]。在我们目前使用的免疫印迹中，p83/100源自PKo菌株（阿氏疏螺旋体），p39（BmpA）和OspC分别源自PKa2菌株（狭义伯氏疏螺旋体）、PKo菌株和PBi菌株（伽氏疏螺旋体）；p41（鞭毛蛋白）内部片段是从PKo菌株和PBi菌株中克隆而来。在本研究中，我们描述了另外两种重组表达的高免疫原性蛋白Osp 7（源自PKo）和p58（源自PBi）的应用。对一组临床明确的血清样本进行了研究，这些样本来自147例莱姆病疏螺旋体病患者和139例对照，之前通过标准化全细胞裂解物免疫印迹进行过检测[Hauser等人（1997年）《临床微生物学杂志》35:1433 - 1444]，在不添加（旧的重组免疫印迹）和添加Osp17和p58（新的重组免疫印迹）的情况下检测重组免疫印迹中的IgG抗体。通过添加Osp17和p58，重组IgG免疫印迹对II期和III期诊断的敏感性可显著提高，且不损失特异性。除了游走性红斑患者的血清外，诊断敏感性与全细胞裂解物IgG免疫印迹相当。重组免疫印迹的主要优点是易于识别诊断条带，而在全细胞裂解物免疫印迹中识别条带则很困难。重组免疫印迹特别适用于需要检测大量血清样本的情况。
