A Serological Diagnostic Immunoassay Test Strip for Leishmaniasis

描述(申请人提供)：利什曼病是印度、非洲、地中海国家、南美国家和德克萨斯州南部的一种地方性疾病。以游客身份前往拉美国家旅游的美国人，以及在伊拉克和阿富汗服役的士兵都感染了这种疾病。聚合酶链式反应和酶免疫吸附试验是诊断本病的常用方法。目前的方法是以实验室为基础的，需要特殊的设备，涉及复杂的程序，并且劳动强度大，耗时长。利什曼病的酶联免疫吸附试验特异性和敏感度都不高，因此有必要开发简便、高灵敏度的利什曼病诊断试验。在这个项目中，智能光学系统公司与耶鲁大学医学院的Diane McMahon-Pratt教授合作，提议开发一种易于使用和高度敏感的横向流动免疫检测试纸，用于准确诊断利什曼病。发展利什曼病免疫分析的传统方法包括使用总的利什曼原虫前鞭毛体作为抗原。在检测中使用这些抗原会降低敏感性，还会导致与相关疾病(如恰加斯病)的交叉反应问题。该项目涉及开发一种更灵敏和更特异的侧向流动免疫分析方法，它将利用高度特异的热休克蛋白作为抗原。这些抗原将与量子点标记一起使用，以实现高灵敏度。在第一阶段，iOS将证明这种检测方法的性能优于目前用于皮肤利什曼病诊断的ELISA试剂盒。使用量子点作为标记，将允许在未来加入更多的抗原，以开发一种多路横向流动免疫分析。与公共卫生相关：利什曼病每年影响数百万人并导致数千人死亡。不幸的是，目前的诊断方法涉及以实验室为基础的劳动密集型程序，而且不够敏感或特异。本申请描述了一种新的方法，用于开发一种简单、现场可用、敏感和高度特异的利什曼病诊断试剂盒。

DESCRIPTION (provided by applicant): Leishmaniasis is a disease endemic to India, Africa, Mediterranean nations, South American countries, and southern Texas. Americans visiting Latin American nations as tourists, and soldiers serving in Iraq and Afghanistan, have contracted the disease. Polymerase chain reaction (PCR) and enzyme immunosorbent assays (ELISA) are commonly used to diagnose this disease. The current methods are laboratory based, require special equipment, involve complex procedures, and are labor intensive and time-consuming. ELISA tests for leishmaniasis are not highly specific and sensitive, so there is an important need to develop easy-to- use and highly sensitive diagnostic tests for leishmaniasis. In this project, Intelligent Optical Systems, in collaboration with Professor Diane McMahon-Pratt of the Yale University School of Medicine, proposes to develop an easy-to-use and highly sensitive lateral flow immuno assay test strip for the accurate diagnosis of leishmaniasis. The conventional approach for developing immunoassays for leishmaniasis involves the use of total Leishmania promastigotes as antigens. Using these antigens in the assay reduces the sensitivity, and also causes a cross reactivity problem with related diseases, such as Chagas disease. This project involves the development of a more sensitive and specific lateral flow immunoassay that will utilize highly specific heat shock proteins as antigens. These antigens will be used, in conjunction with quantum dots labels, to achieve high sensitivity. In Phase I, IOS will demonstrate the superior performance of this assay over the ELISA kits currently used for cutaneous leishmaniasis diagnosis. The use of quantum dots as labels will allow additional antigens to be incorporated, in the future, to develop a multiplexed lateral flow immunoassay. PUBLIC HEALTH RELEVANCE: Leishmaniasis affects millions of people and causes thousands of deaths annually. Unfortunately, current methods of diagnosis involve labor intensive laboratory based procedures, and are not sufficiently sensitive or specific. This application describes a novel method to develop a simple, field-usable, sensitive, and highly specific test kit for leishmaniasis diagnosis.