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在小鼠模型中早期感染期间组织和血浆中伯氏疏螺旋体抗原的检测。

Detection of Borrelia burgdorferi antigens in tissues and plasma during early infection in a mouse model.

机构信息

Paris-Saclay University, CEA, INRAE, Medicines and Healthcare Technologies Department (DMTS), SPI, 91191, Gif-sur-Yvette, France.

出版信息

Sci Rep. 2021 Aug 30;11(1):17368. doi: 10.1038/s41598-021-96861-z.

Abstract

Borrelia burgdorferi is the causative agent of Lyme borreliosis, which is the most common tick-borne human disease in Europe and North America. Currently, the diagnosis of Lyme borreliosis is based on serological tests allowing indirect detection of anti-Borrelia antibodies produced by patients. Their main drawback is a lack of sensitivity in the early phase of disease and an incapacity to prove an active infection. Direct diagnostic tests are clearly needed. The objectives of this study were to produce tools allowing sensitive detection of potential circulating Borrelia antigens and to evaluate them in a mouse model. We focused on two potential early bacterial makers, the highly variable OspC protein and the conserved protein FlaB. High-affinity monoclonal antibodies were produced and used to establish various immunoassays and western blot detection. A very good limit of detection for OspC as low as 17 pg/mL of sample was achieved with SPIE-IA. In infected mice, we were able to measure OspC in plasma with a mean value of 10 ng/mL at 7 days post-inoculation. This result suggests that OspC could be a good blood marker for diagnosis of Lyme borreliosis and that the tools developed during this study could be very useful.

摘要

伯氏疏螺旋体是莱姆病的病原体,是欧洲和北美的最常见的蜱传人类疾病。目前,莱姆病的诊断基于血清学检测,间接检测患者产生的抗伯氏疏螺旋体抗体。其主要缺点是在疾病的早期阶段缺乏敏感性,并且无法证明存在活动性感染。显然需要直接诊断检测。本研究的目的是制备能够灵敏检测潜在循环伯氏疏螺旋体抗原的工具,并在小鼠模型中进行评估。我们集中研究了两种潜在的早期细菌标志物,高度可变的 OspC 蛋白和保守的 FlaB 蛋白。制备了高亲和力的单克隆抗体,并用于建立各种免疫分析和 Western blot 检测。SPIE-IA 检测到的 OspC 检测限非常低,低至 17 pg/mL 的样本。在感染的小鼠中,我们能够在接种后 7 天测量到血浆中的 OspC,平均值为 10 ng/mL。这一结果表明,OspC 可能是莱姆病诊断的一个很好的血液标志物,并且本研究中开发的工具可能非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d24/8405660/4dbfe29a6dab/41598_2021_96861_Fig1_HTML.jpg

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