Simple and rapid POC detection for fungemia

摘要 真菌种类是引起医院内血流和侵袭性感染的最常见原因之一 免疫功能受损的宿主。念珠菌血症的死亡率可能非常高，从20%到40%不等。隐球菌 大约2%-3%的实体器官移植患者受到影响，隐球菌已经成为一种新的 肺炎和播散性真菌病的病因。全球出现毒力多药耐药 金黄色假丝酵母等物种的死亡率据报道高达68%，尤其令人担忧。文化-- 基于检测念珠菌感染的测试速度慢，相对不敏感，而且可能很难区分 在金黄色念珠菌和其他相关念珠菌物种之间。隐球菌侧向流动抗原检测结果较高 敏感度和特异度，但隐球菌抗原检测只能作为一种独立的检测方法，而不是 更大的真菌群的一部分；因此，在没有临床怀疑的情况下，可能会漏掉隐球菌感染。 基于核酸扩增的念珠菌检测方法不具有检测念珠菌所需的灵敏度 直接从患者血液样本感染(不需预培养扩增)，T2念珠菌检测除外 (基于磁共振技术)，其价格高得令人望而却步，速度慢，吞吐量低，而且只有 能够区分有限数量的假丝酵母菌。更广泛的可获得性、更快速的用户 对真菌血流感染进行友好、廉价的检测有可能极大地影响患者 关心和改善感染控制。这项提议将利用多年的成功合作。 在Alland实验室和Cepheid开发的护理点诊断分析之间，包括新的 大血量样本处理和超灵敏DNA序列检测的方法。我们会 利用这一经验创建具有无与伦比的性能的真菌检测分析。我们的具体目标 主要内容有：1)检测发展。2)改进了样品处理，最大限度地增加了测试血量，提高了化验能力 敏感度。3)使用干燥试剂珠进行开发和测试。4)执行分析性研究和临床前研究 为大规模临床试验做准备。

ABSTRACT Fungal species are among the most common causes of nosocomial bloodstream and invasive infections in immunocompromised hosts. Mortality from candidemia can be very high, ranging from 20-40%. Cryptococcus affects approximately 2-3% of solid organ transplant patients and Cryptococcus gattii has emerged as a new cause of pneumonia and disseminated fungal disease. The global emergence of virulent multidrug resistant species such as Candida auris, with a mortality reported to be as high as 68% is particularly worrisome. Culture- based tests to detect Candida infections are slow, relatively insensitive, and may have difficulty distinguishing between C. auris and other related Candida species. Lateral flow antigen tests for Cryptococcus have high sensitivity and specificity but Cryptococcal antigen detection is available only as a standalone assay that is not part of a larger fungal panel; thus, Cryptococcus infections may be missed in the absence of clinical suspicion. Nucleic acid amplification based Candida assays do not have the sensitivity required to detect a Candida infection directly from a patient blood sample (without pre-culture amplification) except for the T2Candida test (based on magnetic resonance technology), which is prohibitively expensive, slow, low throughput and is only able to distinguish among a limited number of Candida species. A more widely available, more rapid and user friendly, and less expensive test for fungal blood stream infections has the potential to greatly impact patient care and improve infection control. This proposal will take advantage of years of successful collaborations between the Alland laboratory and Cepheid developing point of care diagnostic assays, including new approaches to large blood volume sample processing and ultra-sensitive DNA sequence detection. We will leverage this experience to create a fungal detection assay with unparalleled performance. Our specific aims are: 1) Assay development. 2) Improved sample processing to maximize test blood volume and enhance assay sensitivity. 3) Development and testing using dried reagent beads. 4) Perform analytic and pre-clinical studies in preparation for large-scale clinical trials.