Novel non-invasive antigen detection assay for the diagnosis of active visceral leishmaniasis and to monitor the therapy efficacy of this disease

新型非侵入性抗原检测方法，用于诊断活动性内脏利什曼病并监测该疾病的治疗效果

• 批准号: 9343119
• 负责人: Claudia Abeijon
• 金额: $ 61.19万
• 依托单位: DETECTOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9343119
• 关键词: Affect; Age; Antibodies; Antigens; Area; Aspirate substance; Award; Biological Assay; Biological Markers; Bone Marrow; Bone marrow biopsy; Chagas Disease; Child; Clinical; Clinical Research; Communicable Diseases; Communities; Complex; Country; Cutaneous Leishmaniasis; DNA; Data; Detection; Development; Diagnosis; Disease; Eastern Africa; Enzyme-Linked Immunosorbent Assay; Epitopes; Evaluation; Exposure to; Foundations; Freezing; Future; Goals; Gold; HIV; Human; India; Individual; Infection; Leishmania; Leishmania donovani; Leishmania infantum; Liver; Mass Spectrum Analysis; Measures; Monitor; Morbidity - disease rate; Non-Visceral; Parasite resistance; Parasites; Patients; Pharmaceutical Preparations; Phase; Production; Proteins; Publishing; Resources; Risk; Sampling; Schistosomiasis; Sensitivity and Specificity; Serum; Small Business Innovation Research Grant; Solid; South America; Southern Europe; Specificity; Spleen; Standardization; System; Testing; Therapeutic; Toxic effect; Treatment Efficacy; Tuberculosis; Urine; Vaccines; Validation; Visceral Leishmaniasis; Work; accurate diagnosis; base; biomarker development; biomarker selection; co-infection; commercialization; cost; diagnosis standard; disease diagnosis; killings; lymph nodes; mortality; novel; polyclonal antibody; protein biomarkers; tool

ABSTRACT The overall goal of this Phase II project is to validate novel leishmanial protein biomarkers for development of a non-invasive urine-based assay to diagnose active visceral leishmaniasis (VL) and to monitor the therapeutic efficacy of this serious disease. VL is endemic in 47 countries, affects 500,000 people a year and kills more than 50,000, 70% of them children under age 15. VL, also known as kala-azar, is caused by parasites of the Leishmania donovani complex: L. donovani and L. archibaldi in the Old World (primarily India and South Eastern Africa), and L. infantum in the New World (Southern Europe and South America). Global VL morbidity and mortality in many parts of the world are increasing due to co-infection with human immunodeficiency virus. Although VL is usually fatal if not treated promptly, the effective drugs are toxic, expensive and difficult to administer, and untreated people with VL are reservoirs of infection who put others in their communities at risk. The gold standard for diagnosis is observation of the parasites (or detection of parasite DNA) in spleen, liver, lymph node or bone marrow aspirates; serum tests measure anti- parasite antibodies, which cannot distinguish between active VL from either prior exposure to the parasite or subsequent to successful treatment of the disease. There is no vaccine for human VL. The WHO has defined that a key requirement for effective control of this serious worldwide disease is a sensitive non-invasive test that can rapidly and reliably diagnose active VL and identify people who need immediate treatment. Our former published work and the Phase I component of this Phase II application have established the foundation for the development of a simple non-invasive urine test to both diagnose active VL and to monitor the therapy efficacy of this disease. We used mass spectroscopy to initially identify three L. infantum and more recently four new L. donovani proteins excreted in the urine of VL patients. We characterized these antigens, raised polyclonal antibodies against them and developed an antigen detection capture ELISA to diagnose VL. A pilot clinical study defined that the three proteins of L. infantum were present in the urines 19/20 well characterized New World VL patients and in none of more than 60 control urines samples from healthy subjects as well as from non VL patients like suffering from cutaneous leishmaniasis, Chagas’ disease, schistosomiasis and tuberculosis. We are currently validating the recently discovered L. donovani markers. For this Phase II proposal we will use a large panel of urine samples from different areas of the world where VL is endemic to validate all seven discovered L. infantum and L. donovani biomarkers as reliable tools for the accurate diagnosis of active VL and to monitor the therapy of this disease. In addition to a solid preliminary data, a strength of this proposal is our access to a unique resource of urine samples from confirmed VL patients from VL endemic regions around the world.

摘要 该II期项目的总体目标是验证新的利什曼原虫蛋白生物标志物， 开发一种非侵入性的基于尿的检测方法来诊断活动性内脏利什曼病（VL）， 监测这种严重疾病的治疗效果。VL在47个国家流行，影响50万人 每年有超过5万人死亡，其中70%是15岁以下的儿童。VL，也被称为黑热病， 由杜氏利什曼原虫复合体的寄生虫引起：L. donovani和L.阿尔奇巴尔迪在旧世界 （主要是印度和东南非洲）和L.新世界的婴儿（南欧和南欧） 美国）。全球VL发病率和死亡率在世界许多地区由于与VVL的共感染而正在增加。 人体免疫机能丧失病毒。虽然VL如果不及时治疗通常是致命的，但有效的药物是 有毒，昂贵和难以管理，未经治疗的VL患者是感染的宿主， 社区中的其他人处于危险之中。诊断的金标准是观察寄生虫（或 检测寄生虫DNA）在脾脏，肝脏，淋巴结或骨髓穿刺液;血清试验测量抗 - 寄生虫抗体，其不能区分来自先前暴露于寄生虫的活性VL或 在成功治疗疾病之后。没有针对人VL的疫苗。WHO定义 有效控制这一严重的世界性疾病的关键要求是敏感的非侵入性测试， 它可以快速可靠地诊断活动性VL，并确定需要立即治疗的人。 我们以前发表的工作和第二阶段申请的第一阶段组成部分已经建立了 为开发一种简单的非侵入性尿检以诊断活动性VL和监测VL奠定了基础。 本病的治疗效果。我们用质谱初步鉴定了三种L。婴儿和更多 最近，四个新的L。VL患者尿液中排泄的donovani蛋白。我们鉴定了这些抗原， 制备了多克隆抗体，并建立了抗原检测捕获ELISA诊断VL。 一项初步临床研究确定，L.尿中有19/20的婴儿 在新世界VL患者的特征尿中，并且在来自健康人的超过60个对照尿样品中没有一个中， 受试者以及非VL患者如患有皮肤利什曼病，恰加斯病， 血吸虫病和肺结核。我们目前正在验证最近发现的L。Donovani标记。 对于第二阶段的建议，我们将使用来自世界不同地区的大量尿液样本 其中VL是特有的以验证所有七个发现的L。infantum和L. Donovani生物标志物作为可靠的工具 对活动性VL的准确诊断和监测治疗有重要意义。除了固体 初步数据，这一建议的优势是我们获得了独特的尿样资源， 来自世界各地VL流行地区的确诊VL患者。