Biomarker discovery for immunodiagnosis of invasive candidiasis

描述（由申请人提供）：侵袭性念珠菌病（IC）是对危重患者最严重的威胁之一。美国每年有多达6万例病例，在美国，与IC相关的总成本可能高达20 - 40亿美元/年。影响患者预后的最关键因素之一是早期诊断。该项目的目标是一个快速和廉价的免疫测定，将使用血清作为样本，以确定IC的存在。这是诊断念珠菌病的一个未满足的需求，特别是在患者护理点附近的测试。目标人群将是先进的医疗干预和使用强效细胞毒性药物导致念珠菌属深部感染扩散的个体。总体假设是念珠菌属的蛋白抗原。进入血液的浓度可以作为疾病的生物标志物。该方法将是免疫测定两种或更多种不同抗原的存在，所述抗原i）在念珠菌属中共有。导致IC的物质和ii）在感染过程中流入血液。同时检测多种抗原可显著提高预测价值。将使用侧流免疫测定（LFI）和/或酶联免疫测定（ELISA）平台对产品进行检测。到目前为止，我们已经使用了一种新的生物标志物发现过程，称为体内微生物抗原发现（InMAD），以确定5个候选蛋白。特定目标1将重复该实验，并鉴定至少5种其他候选生物标志物（共10种蛋白质）。目标2将使用来自动物模型和人类的IC血清样品确认至少5种蛋白质作为真正的生物标志物。如果I期成功，则II期应用将i）产生针对生物标志物蛋白的单克隆抗体，ii）以LFI或ELISA形式构建原型免疫测定，以及iii）进行用于诊断IC的临床前研究。

DESCRIPTION (provided by applicant): Invasive candidiasis (IC) is one of the most serious threats to patients in a critical setting. With as many as 60,000 cases per year in the U.S., the total cost associated with IC in the U.S. may be as high as $2-4 billion/year. One of the most critical factors influencing patient outcome is early diagnosis. The goal of this project is a rapi and inexpensive immunoassay that will use serum as a sample to identify the presence of IC. This is an unmet need for diagnosis of candidiasis, particularly for tests near the point of patien care. The target population will be individuals for whom advanced medical intervention and use of potent cytotoxic drugs have led to a proliferation of deep-seated infection by Candida spp. The overall hypothesis is that protein antigens of Candida spp. are shed into blood in concentrations that would be biomarkers for disease. The approach will be immunoassay for the presence of two or more distinct antigens that i) are shared across Candida spp. that cause IC and ii) are shed into blood during infection. Simultaneous detection of multiple antigens will markedly enhance the predictive value. The product will be assays with the lateral flow immunoassay (LFI) and/or enzyme-linked immunoassay (ELISA) platforms. To date, we have used a novel biomarker discover process termed In vivo Microbial Antigen Discovery (InMAD) to identify 5 candidate proteins. Specific Aim 1 will repeat this experiment and identify at least 5 additional candidate biomarkers (total of 10 proteins). Aim 2 will confirm at least 5 proteins as genuine biomarkers using IC serum samples from animal models and humans. If Phase I is successful, a Phase II application would i) generate monoclonal antibodies to biomarker proteins, ii) construct prototype immunoassays in LFI or ELISA format and iii) perform pre-clinical studies for diagnosis of IC.