An antigen-detection assay to diagnose Babesia microti infection

诊断田鼠巴贝斯虫感染的抗原检测方法

• 批准号: 10403621
• 负责人: Michel Ledizet
• 金额: $ 69.21万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-10 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403621
• 关键词: Acute respiratory failure; American; Anaplasmosis; Animals; Antibiotics; Antibodies; Antigens; Babesia; Babesia microti; Babesiosis; Biological; Biological Assay; Black-legged Tick; Blood; Blood Transfusion; Blood donor; Blood specimen; Borrelia microti; Centers for Disease Control and Prevention (U.S.); Characteristics; Clinical; Clinical Sensitivity; Complex; Congestive Heart Failure; DNA; Development; Diagnosis; Diagnostic Procedure; Diffuse; Disease; Disease model; Elderly; Enzyme-Linked Immunosorbent Assay; Epitopes; Erythrocytes; Evaluation; Goals; Gold; Health; Human; Immunocompetent; Immunocompromised Host; Immunofluorescence Immunologic; Individual; Industry Standard; Infection; Institute of Medicine (U.S.); Kidney Failure; Laboratories; Lead; Longitudinal prospective study; Lyme Disease; Malaria; Methods; Microscopic; Microscopy; Monitor; Monoclonal Antibodies; Mus; Parasitemia; Parasites; Patients; Performance; Phase; Population; Prospective Studies; Proteins; RNA; Recombinant Proteins; Recurrent disease; Red Cross; Relapse; Reporting; Resolution; Retrospective Studies; Risk; Sampling; Sensitivity and Specificity; Serology; Serum; Signs and Symptoms; Specificity; Statistical Data Interpretation; Symptoms; Techniques; Ticks; Transfusion; Treatment Failure; United States; Vascular blood supply; active method; antibody detection; antigen detection; assay development; base; cohort; cost; design; detection assay; diagnostic assay; experimental study; flu; granulocyte; human disease; human model; immunosuppressed; infection risk; mortality; mouse model; pathogen; rapid test; tick transmission; vector tick

PROJECT SUMMARY Human babesiosis is a malaria-like multisystem disease caused primarily by Babesia microti, an emerging apicomplexan parasite that infects and develops within human erythrocytes. The parasite is transmitted to humans by the tick vector Ixodes scapularis and can also be introduced through blood transfusion. Infection can cause flu-like symptoms, and severe infection can be fatal, in particular in the immunosuppressed and the elderly. Current methods for babesiosis diagnosis include microscopy, PCR, IFA and ELISA-based methods that detect antibodies in serum from patients or donors. Each of these methods has major limitations, such as insufficient sensitivity, high complexity, and low throughput. In addition, results from many of these assays remain positive for months or years after resolution of active infection. In Phase I of this project, we have developed a capture ELISA assay that can detect a protein of the parasite, BmGPI12/BmSAI. We have identified pairs of monoclonal antibodies that can be used to capture BmGPI12 in mouse and human samples. Our preliminary results demonstrate that this assay has an excellent sensitivity. Using samples collected from infected mice, a well-established model of human disease, we demonstrated that following successful treatment, our assay gives negative results while PCR and serology remain positive. Thus, our assay may be useful in distinguishing active infection from past exposure. In Phase II of this project, we will further develop this assay with the goal of applying for FDA clearance. We will analyze several hundred animal and human samples to establish important characteristics of the assay including sensitivity and specificity. These Phase II experiments are designed to give us a full understanding of the potential biological indications and commercial usefulness of the assay.

项目摘要 人类巴布西病是一种类似疟疾的多系统疾病 在人类红细胞中感染和发展的Apicomplexan寄生虫。寄生虫传播到 tick矢量ixodes肩cap骨的人类，也可以通过输血引入。感染 会引起流感样症状，严重感染可能是致命的，特别是在免疫抑制和 老年。当前的Babesiosis诊断方法包括显微镜，PCR，IFA和基于ELISA的方法 该检测患者或供体的血清中的抗体。这些方法中的每一个都有主要的局限性，例如 灵敏度不足，高复杂性和低吞吐量。此外，许多这些测定法 分辨活性感染后几个月或几年保持阳性。 在该项目的第一阶段，我们开发了一种捕获ELISA分析，可以检测寄生虫的蛋白质， BMGPI12/BMSAI。我们已经确定了可用于捕获BMGPI12的单克隆抗体对 小鼠和人类样品。我们的初步结果表明，该测定法具有出色的灵敏度。 使用从受感染小鼠的样品（一种已建立的人类疾病模型）中收集的样品，我们证明了 成功治疗后，我们的测定在PCR和血清学保持阳性的同时产生负面结果。 因此，我们的测定可能有助于区分活性感染和过去的暴露。 在该项目的第二阶段中，我们将进一步开发该测定法，以申请FDA清除率。我们 将分析数百个动物和人类样品以建立测定的重要特征 包括灵敏度和特异性。这些第二阶段的实验旨在使我们充分理解 测定的潜在生物学适应症和商业实用性。

项目成果

Babesia BdFE1 esterase is required for the anti-parasitic activity of the ACE inhibitor fosinopril.

• DOI: 10.1016/j.jbc.2023.105313
• 发表时间: 2023-11
• 期刊: The Journal of biological chemistry
• 作者: Vydyam P;Choi JY;Gihaz S;Chand M;Gewirtz M;Thekkiniath J;Lonardi S;Gennaro JC;Ben Mamoun C
• 通讯作者: Ben Mamoun C

Babesia duncani multi-omics identifies virulence factors and drug targets.

• DOI: 10.1038/s41564-023-01360-8
• 发表时间: 2023-05
• 期刊: NATURE MICROBIOLOGY
• 影响因子: 28.3
• 作者: Singh, Pallavi;Lonardi, Stefano;Liang, Qihua;Vydyam, Pratap;Khabirova, Eleonora;Fang, Tiffany;Gihaz, Shalev;Thekkiniath, Jose;Munshi, Muhammad;Abel, Steven;Ciampossin, Loic;Batugedara, Gayani;Gupta, Mohit;Lu, Xueqing Maggie;Lenz, Todd;Chakravarty, Sakshar;Cornillot, Emmanuel;Hu, Yangyang;Ma, Wenxiu;Gonzalez, Luis Miguel;Sanchez, Sergio;Estrada, Karel;Sanchez-Flores, Alejandro;Montero, Estrella;Harb, Omar S.;Le Roch, Karine G.;Ben Mamoun, Choukri
• 通讯作者: Ben Mamoun, Choukri

An Alternative Culture Medium for Continuous In Vitro Propagation of the Human Pathogen Babesia duncani in Human Erythrocytes.

• DOI: 10.3390/pathogens11050599
• 发表时间: 2022-05-20
• 期刊: Pathogens (Basel, Switzerland)

Tafenoquine-Atovaquone Combination Achieves Radical Cure and Confers Sterile Immunity in Experimental Models of Human Babesiosis.

他非诺喹-阿托伐醌组合在人类巴贝虫病实验模型中实现了根治并赋予了无菌免疫力。

• DOI: 10.1093/infdis/jiad315
• 发表时间: 2024
• 期刊: The Journal of infectious diseases
• 作者: Vydyam,Pratap;Pal,AnasuyaC;Renard,Isaline;Chand,Meenal;Kumari,Vandana;Gennaro,JosephC;Mamoun,ChoukriBen
• 通讯作者: Mamoun,ChoukriBen