Quantitation and Genotyping of HCV RNA by Time Resolved Lanthanide Luminescence

描述（由申请人提供）：丙型肝炎病毒（HCV）是一种约9.4 kb的单链RNA病毒，估计影响全球2亿人。在美国，据估计有410万人感染，HCV占慢性肝炎的60-70%和肝硬化、终末期肝病和肝细胞癌的50%，每年造成估计12，000人死亡。从1994年到2007年，每年因HCV住院的人数增加了10.3倍，而HCV单一感染患者的总医疗支出从6.27亿美元增加到69亿美元，HIV/HCV合并感染患者的总医疗支出从6300万美元增加到6.55亿美元。在美国，大约75%的HCV感染者不知道自己的病情，并且患肝硬化和肝癌晚期并发症的风险增加。机构外的患者通道 由于检测和治疗费用高昂，医疗保健系统受到限制，特别是对于没有保险和保险不足的人-只有54%的HCV治疗候选人有任何类型的保险。美国退伍军人管理局医院的患者，城市美洲原住民人口和少数民族的慢性HCV感染和并发症的发病率不成比例地高。 目前的标准治疗组合抗病毒治疗（利巴韦林/聚乙二醇干扰素）根除了许多患者的HCV，而新FDA批准的HCV特异性治疗大大增加了积极治疗结果的可能性和速度。当前和新兴治疗方案成功实施的先决条件是基因分型和病毒载量测量，这两者都有助于确定治疗过程和持续时间。基线HCV RNA水平的测定对于监测患者对治疗的反应至关重要，最近的证据表明，基于初始病毒载量和病毒学反应的个性化治疗可提高成功率。随着HCV标准治疗的发展，识别HCV感染者变得更加重要，包括重新推动筛查计划以识别和治疗未确诊的患者，以及快速护理点检测。 该提案开发了EOSCAPE-CLEF（螯合镧系元素发射指纹），这是一种低成本的护理点（POC）替代传统HCV RNA检测，将通过信号放大进行定量检测与敏感和特异性基因分型相结合，使用基于与严格分子信标式探针缀合的单个螯合镧系元素对的特征光发射谱的开创性信号处理算法。预计该系统将把定量和基因分型的成本从400美元降低到35美元以下，同时将周转时间缩短到30分钟。该测定在基于申请人组织的整体狭缝毛细管阵列流体微致动器（mSCAFA）技术的一次性封闭盒中运行。 公共卫生相关性：丙型肝炎病毒（HCV）感染占肝硬化，终末期肝病和肝癌的50%，在美国每年估计造成10，000至12，000人死亡，是全球的许多倍。HCV的成功治疗取决于亚型鉴定和通过专门的类型诊断测试频繁测量血液中的病毒量。该提案将开发一种创新设备，用于快速和廉价的全血HCV联合定量和基因分型，这将改善患者结局并降低国内和全球HCV患者护理的成本。

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is a single-stranded RNA virus of roughly 9.4 kb estimated to affect 200 million people worldwide. In the U.S., where it is estimated that 4.1 million are infected, HCV accounts for 60-70% of chronic hepatitis and 50% of cirrhosis, end-stage liver disease, and hepatocellular carcinoma, causing an estimated 12,000 deaths annually. From 1994 to 2007, annual hospitalizations for HCV rose 10.3-fold, while total healthcare spending increased from $627 million to $6.9 billion in HCV mono-infected patients and from $63 million to $655 million in HIV/HCV co-infected patients. Approximately 75% of HCV-infected people in the U.S. are unaware of their condition and are at increased risk for late-stage complications of cirrhosis and liver cancer. Patient access outside of institutional healthcare systems is limited by the high cost of testing and treatment, particularly for the uninsured and underinsured - only 54% of HCV treatment candidates have any type of insurance coverage. Patients at Veterans Administration hospitals, urban Native American populations, and ethnic minorities in the U.S. have disproportionately high rates of chronic HCV infection and complications. Current standard-of-care combination antiviral therapy (ribavirin/peg-interferon) eradicates HCV in many patients, while newly FDA-approved HCV-specific therapies greatly increase the likelihood and speed of positive treatment outcomes. Prerequisites for successful administration of current and emerging therapeutic regimens are genotyping and measurement of viral loads, both of which help determine treatment course and duration. Determination of baseline HCV RNA levels is critical for monitoring patient response to therapy, and recent evidence suggests that personalizing treatment based on initial viral load and virological response increases success rates. As the HCV standard of care evolves, identifying people infected with HCV takes on even greater importance, including a renewed push for screening programs to identify and treat undiagnosed patients, as well as for rapid point of care testing. This proposal develops the EOSCAPE-CLEF (Chelated Lanthanide Emission Fingerprinting), a low-cost, point of care (POC) alternative to conventional HCV RNA testing, combining quantitative detection by signal amplification with sensitive and specific genotyping using a pioneering signal processing algorithm based on the characteristic light emission profiles of individual chelate-lanthanide pairs conjugated to a stringent molecular beacon-style probe. The enclosed-cartridge format system is anticipated to reduce costs from a combined $400 for quantitation and genotyping to less than $35 while also reducing the turnaround time to 30 minutes. The assay runs in a single-use enclosed cartridge based on the Applicant Organization's monolithic slit capillary array fluidic microactuator (mSCAFA) technology. PUBLIC HEALTH RELEVANCE: Infection with the Hepatitis C virus (HCV) accounts for up to 50 percent of cirrhosis, end-stage liver disease, and liver cancer and causes an estimated 10,000 to 12,000 deaths annually in the United States and many times more than that globally. Successful treatment of HCV depends upon subtype identification and frequent measurement of the amount of virus in the blood through a specialized type diagnostic testing. This proposal will develop an innovative device for fast and inexpensive combined quantitation and genotyping of HCV from whole blood which will improve patient outcomes and reduce costs for HCV patient care both domestically and globally.