Semiconductor Electronic Label-Free Assay (SELFA) for Diagnosing Heart Disease

简介(申请人提供)：心血管疾病是全球发病率和死亡率的主要原因。仅在美国和欧洲，每年就有大约1500万名急性冠脉综合征患者被送往急诊科。这些患者立即通过心电图和血液样本中的生物标记物检测来筛查潜在的急性心肌梗死(AMI)。由于心肌肌钙蛋白I(CTnI)的高度特异性和在事件发生后数小时内血液中的浓度升高，cTnI已成为诊断急性心肌梗死的最可靠的生物标志物之一，当cTnI浓度低于0.10 ng/mL被认为是阴性结果，而cTnI浓度大于或等于0.10 ng/mL被认为是阳性结果。此外，最近的研究表明，cTnI也可以用于急性心肌梗死的危险分层。然而，对于这一预期用途，需要使用超灵敏分析来检测血液样本中略有升高的cTnI水平。 目前，有两种方法可用于测定患者样本中cTnI的浓度：基于实验室的检测方法和护理点(POC)检测方法。虽然基于实验室的分析更敏感，但它们的周转时间(TAT)较长，通常为45分钟至1小时，这对于诊断速度至关重要的急性心肌梗死等应用程序并不理想。另一方面，虽然目前的POC检测已经大大改进了10-15分钟的TAT，并且可以在床边实施，但它们的灵敏度较低，因此被发现不如基于实验室的检测那么可靠。此外，即使是目前基于实验室的分析方法，在量化患者样本中的cTnI以帮助进行急性心肌梗死风险分层方面也没有必要的分析灵敏度。 我们SELFA Inc.的目标是开发一种新的、敏感的无标记POC方法，用于快速、可靠和准确地检测患者血液样本中的cTnI，有助于急性心肌梗死的诊断和风险分层。这项检测将基于一种新的半导体纳米线场效应晶体管技术，该技术利用抗cTnI抗体来定量患者样本中的cTnI。在初步研究中，我们已经证明，与目前基于实验室的检测方法相比，我们在检测cTnI浓度方面的灵敏度提高了1000倍。我们的新检测方法将包括一次性使用的试纸，以及易于使用和便携式的阅读器。在这项快速通道计划的第一阶段，我们将(A)检查各种抗cTnI抗体-cTnI组合，以最大限度地提高我们生物传感器的灵敏度，以及(B)开发和优化第0代实验室级SELFA测试试条原型，并验证其分析和临床性能。在第一阶段成功完成后，在本提案的第二阶段，我们将(C)开发和优化第一代商业级SELFA测试条和便携式阅读器设备，(D)建立和评估用于高通量校准测试条的稳健战略，以及最后(F)验证商业级测试条的分析和临床性能，并准备和启动FDA监管批准程序。

DESCRIPTION (provided by applicant): Cardiovascular disease is the leading cause of morbidity and mortality worldwide. In United States and Europe alone, about 15 million patients are annually admitted to emergency departments with acute coronary syndrome. These patients are immediately screened for potential acute myocardial infarction (AMI) via electrocardiogram and biomarker detection in blood samples. Due to its high specificity and elevated concentrations in blood for hours past an incident, cardiac troponin I (cTnI) has become one of the most reliable biomarkers for diagnosing AMI where a cTnI concentration of less than 0.10 ng/mL is considered a negative result, while a cTnI concentration of 0.10 ng/mL or greater is considered a positive result. Furthermore, recent studies have shown that cTnI could also be used for AMI risk stratification. However, for this intended use, ultra-sensitive assays are required to detect slightly elevated levels of cTnI in blood samples. Currently there are two methods available for determining the concentration of cTnI in patient samples: lab-based assays and point-of-care (POC) assays. While lab-based assays are more sensitive, they have a longer turnaround time (TAT) of typically 45 minutes to 1 hour, which is not ideal for application such as diagnosing AMI where speed is crucial. On the other hand, while current POC assays have much improved TAT of 10-15 minutes and could be implemented at bed-side, they have inferior sensitivity, and hence found to not be as reliable as lab-based assays. Additionally, even the current lab-based assays do not have the necessary analytical sensitivity in quantifying cTnI in patient samples to aid in AMI risk stratification. We at Selfa Inc. aim to develop a novel and sensitive label-free POC assay for rapid, reliable, and accurate detection of cTnI in patient blood samples that could aid in diagnosing and risk stratification of AMI. This assay will be based on a novel semiconductor nanowire field-effect transistor technology that utilizes anti-cTnI antibodies to quantify cTnI in a patient sample. In or preliminary studies, we have demonstrated a 1000-fold improved sensitivity in detecting cTnI concentrations compared to current lab-based assays. Our novel assay will consist of disposable, one-time-use test strips, and an easy-to-use and portable reader. In Phase I of this Fast-Track proposal, we will (a) examine various anti-cTnI antibody-cTnI combinations to maximize the sensitivity of our biosensor, and (b) develop and optimize the 0th generation, lab-grade SELFA test strips prototype and validate their analytical and clinical performance. Upon successful completion of Phase I, in Phase II of this proposal, we will (c) develop and optimize the 1st generation, commercial-grade SELFA test strips and portable reader device, (d) establish and evaluate a robust strategy for high throughput calibration of the test strips, and finally (f) validate the analytical and clinical performance of the commercial-grade test strips an reader, and prepare and initial the FDA regulatory approval process.