Biomarker discovery for immunodiagnosis of invasive candidiasis

描述(由申请人提供)：侵袭性念珠菌病(IC)是危重患者面临的最严重威胁之一。在美国，每年有多达6万个病例，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。这个项目的目标是一种快速且廉价的免疫分析方法，它将使用血清作为样本来识别IC的存在。这是对念珠菌病诊断的一种未得到满足的需求，特别是在患者护理点附近的测试中。目标人群将是先进的医疗干预和强有力的细胞毒性药物的使用导致念珠菌深层感染扩散的个人。总的假设是假丝酵母菌的蛋白质抗原。进入血液的浓度将成为疾病的生物标志物。该方法将通过免疫分析检测是否存在两个或更多不同的抗原，这些抗原在念珠菌中是共有的。导致IC和II)的病毒在感染过程中被排入血液。同时检测多种抗原将显著提高其预测价值。该产品将通过侧向流动免疫分析(LFI)和/或酶联免疫分析(ELISA)平台进行检测。到目前为止，我们已经使用了一种被称为体内微生物抗原发现(InMAD)的新的生物标记物发现过程来鉴定5个候选蛋白质。特殊目标1将重复这个实验，并确定至少5个额外的候选生物标记物(总共10个蛋白质)。目标2将使用动物模型和人类的IC血清样本确认至少5种蛋白质为真正的生物标志物。如果第一阶段成功，第二阶段的应用将：i)产生针对生物标记蛋白的单抗；ii)构建LFI或EL ISA格式的免疫分析原型；iii)进行诊断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.