Department of Molecular Biotechnology and Microbiology, Faculty of Chemistry, University of Gdańsk Technology, 80-233 Gdańsk, Poland.
SaBio, Instituto de Investigación en Recursos Cinegéticos IREC-CSIC-UCLM-JCCM, 13005 Ciudad Real, Spain.
ACS Infect Dis. 2023 Nov 10;9(11):2160-2172. doi: 10.1021/acsinfecdis.3c00258. Epub 2023 Oct 6.
Lyme disease is a tick-borne zoonosis caused by Gram-negative bacteria belonging to the sensu lato (s.l.) group. In this study, IgM- and IgG-specific linear epitopes of two sensu stricto (s.s.) antigens BmpA and BBK32 were mapped using a polypeptide array. Subsequently, two chimeric proteins BmpA-BBK32-M and BmpA-BBK32-G were designed to validate the construction of chimeras using the identified epitopes for the detection of IgM and IgG, respectively, by ELISA. IgG-ELISA based on the BmpA-BBK32-G antigen showed 71% sensitivity and 95% specificity, whereas a slightly lower diagnostic utility was obtained for IgM-ELISA based on BmpA-BBK32-M, where the sensitivity was also 71% but the specificity decreased to 89%. The reactivity of chimeric proteins with nondedicated antibodies was much lower. These results suggest that the identified epitopes may be useful in the design of new forms of antigens to increase the effectiveness of Lyme disease serodiagnosis. It has also been proven that appropriate selection of epitopes enables the construction of chimeric proteins exhibiting reactivity with a specific antibody isotype.
莱姆病是一种由革兰氏阴性细菌引起的蜱传动物病，属于广义（s.l.）组。在这项研究中，使用多肽阵列对两种狭义（s.s.）抗原 BmpA 和 BBK32 的 IgM 和 IgG 特异性线性表位进行了作图。随后，设计了两种嵌合蛋白 BmpA-BBK32-M 和 BmpA-BBK32-G，分别使用鉴定的表位通过 ELISA 验证用于检测 IgM 和 IgG 的嵌合体构建。基于 BmpA-BBK32-G 抗原的 IgG-ELISA 显示出 71%的敏感性和 95%的特异性，而基于 BmpA-BBK32-M 的 IgM-ELISA 的诊断效果略低，其敏感性也为 71%，但特异性降低至 89%。嵌合蛋白与非专用抗体的反应性要低得多。这些结果表明，鉴定的表位可用于设计新形式的抗原以提高莱姆病血清学诊断的有效性。还证明了适当选择表位可以构建具有特定抗体同种型反应性的嵌合蛋白。
