Grąźlewska Weronika, Ferra Bartłomiej, Rudzińska Monika, Holec-Gąsior Lucyna
Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, Gdańsk University of Technology, 80-233 Gdańsk, Poland.
Department of Tropical Medicine and Epidemiology, Institute of Maritime and Tropical Medicine, Medical University of Gdańsk, 81-519 Gdynia, Poland.
Pathogens. 2021 Jun 18;10(6):767. doi: 10.3390/pathogens10060767.
Currently, the diagnosis of Lyme disease is based mostly on two-tiered serologic testing. In the new generation of immunoenzymatic assays, antigens comprise whole-cell lysates of members of the sensu lato (s.l.) species complex, with the addition of selected recombinant proteins. Due to the high diversity of members of the s.l. genospecies and the low degree of conservation among the amino acid sequences of their proteins, serodiagnostic methods currently in use are not sufficient for the correct diagnosis of borreliosis. Two divalent chimeric proteins (BmpA-BBK32 and BmpA-BBA64) were expressed in . Following purification by one-step metal-affinity chromatography, preparations were obtained containing milligram levels of chimeric protein exhibiting electrophoretic purity in excess of 98%. Reactivity of the new chimeric proteins with specific human IgG antibodies was preliminarily determined by Western blot. For this purpose, 20 negative sera and 20 positive sera was used. The new chimeric proteins were highly reactive with IgG antibodies contained in the serum of patients suffering from borreliosis. Moreover, no immunoreactivity of chimeric proteins was observed with antibodies in the sera of healthy people. These promising results suggest that new chimeric proteins have the potential to discriminate between positive and negative sera.
目前,莱姆病的诊断主要基于两层血清学检测。在新一代免疫酶测定中,抗原包括狭义(s.l.)种复合物成员的全细胞裂解物,并添加了选定的重组蛋白。由于狭义基因种成员的高度多样性及其蛋白质氨基酸序列之间的低保守程度,目前使用的血清诊断方法不足以正确诊断疏螺旋体病。两种二价嵌合蛋白(BmpA - BBK32和BmpA - BBA64)在……中表达。通过一步金属亲和层析纯化后,获得了含有毫克级嵌合蛋白的制剂,其电泳纯度超过98%。通过蛋白质印迹初步确定了新嵌合蛋白与特异性人IgG抗体的反应性。为此,使用了20份阴性血清和20份阳性血清。新嵌合蛋白与莱姆病患者血清中所含的IgG抗体具有高度反应性。此外,在健康人血清中未观察到嵌合蛋白与抗体的免疫反应性。这些有前景的结果表明,新嵌合蛋白有区分阳性和阴性血清的潜力。