Immunoassay for early diagnosis of mucormycosis

毛霉菌病是最致命的侵袭性真菌感染之一。毛霉菌病最常发生在 接受化疗的恶性血液病患者，接受造血干细胞移植的患者 干细胞移植或糖尿病患者。改善诊断是最常见的未满足 需要管理毛霉菌病患者。延迟治疗会增加死亡率， 已经高达47%到83%。与其他侵袭性真菌感染不同， 毛霉菌病的诊断然而，在我们的初步研究中， 所产生的与毛霉目的细胞壁岩藻甘露聚糖具有高反应性。第一代免疫分析 从mAb 2DA6产生的，在血清、尿液、支气管肺泡液和感染组织中发现了岩甘聚糖， 临床相关的小鼠毛霉菌病模型以及来自人类毛霉菌病病例的血浆和尿液中。 目标是一种免疫测定法，使用血浆或尿液快速诊断早期毛霉菌病。目标 人群是指糖尿病或使用强效免疫抑制药物导致 毛霉菌病的发病率急剧上升。该方法是一种免疫测定法， 葡甘露聚糖是一种细胞壁碳水化合物，由许多产生毛霉病的接合菌所共有。的 该产品将是一种快速、廉价和易于使用的侧流免疫测定法（LFIA）。 有四个具体目标。目的1将证实mAb 2DA 6在各种接合菌中的反应性， 产生毛霉菌病。目的2优化免疫分析法检测岩甘聚糖的样品前处理方法。 目标3将确定最佳检测灵敏度和特异性所需的免疫检测限， 从毛霉菌病或曲霉病患者中前瞻性采集的充分表征的样本，以及 对照组，而不是受感染的患者。目标4将构建一个先进的原型LFIA，满足早期检测截止值 和毛霉菌病的特异性诊断。 这项提案解决了一个关键的未满足的需要，越来越常见和致命的机会性真菌 感染该测试的临床用途和市场规模将与目前可用的免疫测定相同， 侵袭性曲霉病的诊断，因为毛霉菌病是对患者的鉴别诊断 疑似侵袭性曲霉病这项研究是有史以来第一次证明和利用一种分泌的真菌 用于诊断毛霉菌病的生物标志物。项目的初步结果，加上广泛接受的临床使用 以及其它靶向细胞壁甘露聚糖的免疫测定用于诊断侵袭性真菌感染的商业成功 疾病，使商业成功的可能性非常高。

Mucormycosis is one of the deadliest of the invasive fungal infections. Mucormycosis occurs most often in patients with hematological malignancies undergoing chemotherapy, patients who have received hematopoietic stem cell transplants or patients with diabetes mellitus. Improved diagnosis is the most frequently noted unmet need for management of the mucormycosis patient. Delays in treatment increase the mortality rate from an already high 47% to 83%. Unlike other invasive fungal infections, there has been no known fungal biomarker for diagnosis of mucormycosis. However, in our preliminary studies, a monoclonal antibody (mAb 2DA6) was produced that has high reactivity with cell wall fucomannan of the Mucorales. A first-generation immunoassay produced from mAb 2DA6 found fucomannan in serum, urine, broncho alveolar fluid and infected tissue from clinically relevant mouse models of mucormycosis and in plasma and urine from human cases of mucormycosis. The goal is an immunoassay that uses plasma or urine to rapidly diagnose early-stage mucormycosis. The target population is individuals for whom diabetes mellitus or use of potent immunosuppressive drugs has led to a dramatic increase in the occurrence of mucormycosis. The approach is an immunoassay for the presence of fucomannan, a cell wall carbohydrate that is shared by the many Zygomycetes that produce mucormycosis. The product will be a lateral flow immunoassay (LFIA) that is rapid, inexpensive, and easy to use. There are four Specific Aims. Aim 1 will confirm reactivity of mAb 2DA6 across the various Zygomycetes that produce mucormycosis. Aim 2 will optimize specimen treatment for detection of fucomannan by immunoassay. Aim 3 will identify the immunoassay limit of detection needed for optimal assay sensitivity and specificity using well characterized, prospectively collected samples from patients with mucormycosis or aspergillosis, as well as control, not infected patients. Aim 4 will construct an advanced prototype LFIA that meets assay cutoffs for early and specific diagnosis of mucormycosis. This proposal addresses a critical unmet need for an increasingly common and fatal opportunistic fungal infection. Clinical use and market size for the test would be identical to currently available immunoassays for diagnosis of invasive aspergillosis because mucormycosis is on the differential diagnosis for patients with suspected invasive aspergillosis. The study is the first ever to demonstrate and exploit a secreted fungal biomarker for diagnosis of mucormycosis. Project preliminary results, coupled with the well-accepted clinical use and commercial success of other immunoassays that target cell wall mannans for diagnosis of invasive fungal disease, make the probability for commercial success very high.