A Serological Diagnostic Immunoassay Test Strip for Leishmaniasis

描述（由申请人提供）：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病。在拉丁美洲国家旅游的美国人，以及在伊拉克和阿富汗服役的士兵都感染了这种疾病。聚合酶链反应（PCR）和酶免疫吸附试验（ELISA）是诊断本病的常用方法。目前的方法是基于实验室的，需要特殊的设备，涉及复杂的程序，劳动密集型和耗时。ELISA法检测利什曼病的特异性和敏感性不高，因此有必要开发易于使用、灵敏度高的利什曼病诊断检测方法。在这个项目中，Intelligent Optical Systems与耶鲁大学医学院的Diane McMahon-Pratt教授合作，建议开发一种易于使用且高度敏感的横向流动免疫测定试条，用于准确诊断利什曼病。开发利什曼病免疫测定法的常规方法包括使用利什曼原虫总原鞭毛体作为抗原。在检测中使用这些抗原降低了敏感性，并且还导致了与诸如恰加斯病等相关疾病的交叉反应性问题。该项目涉及开发一种更敏感和特异性的侧流免疫分析法，该方法将利用高度特异性的热休克蛋白作为抗原。这些抗原将与量子点标签一起使用，以实现高灵敏度。在I期试验中，IOS将证明该检测方法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额外的抗原，以开发多路侧流免疫测定。公共卫生相关性：利什曼病每年影响数百万人，造成数千人死亡。不幸的是，目前的诊断方法涉及劳动密集型的实验室程序，不够敏感或特异性。本应用程序描述了一种开发简单，现场可用，敏感和高度特异性的利什曼病诊断检测试剂盒的新方法。

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.