Point-of-Care Microfluidic Biochip for Biomarkers Monitoring for Contributing in Early Sepsis Diagnosis

首席调查员/项目主任(最后、第一、中间)：拉希德·巴希尔 项目摘要：脓毒症是一种威胁生命的器官功能障碍，由宿主对感染的反应失调引起(脓毒症-- 3定义)，是导致死亡的主要原因，也是医院最昂贵的条件。每年，有3000万人受到影响 在世界范围内，美国每年至少有170万成年人患上脓毒症(近27万人死亡)，成本为240亿美元。 被诊断为脓毒症且没有持续的器官衰竭迹象的患者死亡的可能性约为15%-30%。然而， 严重败血症或感染性休克患者的死亡率可增加到40%-60%。在医院死亡的患者中每三人中就有一人 有败血症。死亡率上升的一个主要因素是无法准确和快速地诊断潜在的 败血症患者。同样，败血症是再次住院的主要原因(比因心脏病住院的比例更高 美国的心脏病发作、心力衰竭、慢性阻塞性肺病和肺炎)。急诊室和重症监护室依赖于极不特定的监测 参数(例如发烧、低血压、心率加快)启动临床诊断并开始治疗。这些 粗略的指标会导致医生将早期脓毒症与其他几种疾病混淆。一例早期发病的阳性诊断 脓毒症是严重的，因为死亡率随着治疗的延迟而增加。据报道，存活率下降了7.6%。 每一小时没有给予适当的抗生素，这些延误增加了不必要的医院费用。超过了 在过去的30年里，临床使用了不同的标准，如SIRS，LODS和SOFA或QSOFA作为筛查工具来评估 潜在败血症患者器官功能障碍的严重程度。这些标准的共同因素是非特异性和 非常高的假阳性率。对于阳性标准的患者，最终的诊断测试是血液培养，这可能需要 至5天为阴性结果。同样，血培养有很高的假阴性率(60%)，不适用于 挑剔的病原体，如肺炎衣原体。更重要的是，血液培养不能成为黄金标准。 用于脓毒症的诊断。这项技术只检测血液中细菌的存在(菌血症)，而不是 必然表明有病。许多非细菌性感染也可能导致危及生命的败血症。为了改善这一状况 脓毒症诊断的准确性和敏感性，脓毒症-3的定义强调了对这两种病原体的要求 检测和有关患者免疫系统的个性化状态的信息。因此，我们建议将重点放在 我们在监测这种免疫反应的选择性生物标记物方面所做的努力。然而，不是单一的，甚至不是组合的 生物标志物已被证实可用于脓毒症的诊断。因为没有一种单一的生物标志物可以预测脓毒症， 我们建议开发一种护理点式微流控生物芯片，用于测量细胞表面和血浆蛋白质生物标记物 将有助于脓毒症的早期诊断。微流控生物芯片将提供完整的白血球 白细胞计数(WBC)以及中性粒细胞(NCD64)、降钙素原(PCT)、C反应蛋白CD64表达的定量 (CRP)和IL-6(IL-6)。多项研究表明，这些生物标志物对脓毒症具有很高的敏感性。这个 拟议的设备将首次结合细胞表面蛋白和血浆蛋白生物标记物的分析 同样的血样。这样的设备，结合医院常规进行的测试，可以显著地 加快脓毒症的诊断和临床决策，为患者提供正确的治疗。

Principal Investigator/Program Director (Last, first, middle): Bashir, Rashid Project Summary: Sepsis, a life-threatening organ dysfunction caused by a dysregulated host response to infection (Sepsis- 3 definition), is the leading cause of death and most expensive condition in hospitals. Annually, > 30 million people affected worldwide, with at least 1.7 million adults developing sepsis (nearly 270K die) at a cost of $24 billion per year in the U.S. Patients diagnosed with sepsis and no ongoing sign of organ failure have about a 15-30% chance of death. However, the mortality rate can increase up to 40-60% for severe sepsis or septic shock patients. One in three patients who die in a hospital have sepsis. One major factor in these rising mortality rates is the inability to accurately and quickly diagnose potentially septic patients. Likewise, sepsis is a leading cause of hospital readmission (higher proportion than hospitalizations for heart attack, heart failure, COPD, and pneumonia in the U.S.). EDs and ICUs rely on monitoring extremely non-specific parameters (e.g. fever, low blood pressure, increased heart rate) to initiate a clinical diagnosis and begin treatment. These crude indicators cause doctors to mistake early stage sepsis with several other diseases. A positive diagnose of early onset sepsis is critical because mortality increases with delays in treatment. Survival rates have been reported to drop by 7.6% every hour that the proper antibiotics are not administered, and these delays compound unnecessary hospital costs. Over the last 30 years, clinics have used different criteria such as SIRS, LODS and SOFA or qSOFA as screening tools to assess the severity of organ dysfunction in a potentially septic patient. Common factors among these criteria are non-specificity and very high false positive rates. For patients with positive criteria, the final diagnostic test is a blood culture that may take up to 5 day for a negative result. Likewise, blood culture has a very high false negative rate (> 60%) and does not work for fastidious pathogens such as Chlamydia pneumoniae. More importantly, blood culture cannot be a gold standard method for sepsis diagnosis. This technique only detects the presence of bacteria in the bloodstream (bacteremia), which does not necessarily indicate illness. Many non-bacteremic infections can also cause life-threatening sepsis. In order to improve the accuracy and sensitivity of sepsis diagnosis, the Sepsis-3 definition underscores the requirements for both pathogen detection and information about the personalized state of the immune system of the patient. Therefore, we propose to focus our efforts on monitoring selective biomarkers of this immune response. However, no single, or even a combination of biomarkers has been validated for the diagnosis of sepsis. Because no single biomarker is specific enough to predict sepsis, we propose to develop a point-of-care microfluidic biochip for measuring cell-surface and plasma-proteins biomarkers that will be used for contributing in early sepsis diagnosis. The microfluidic biochip will provide a complete white blood cell count (WBC), as well as quantification of CD64 expression on neutrophil (nCD64), procalcitonin (PCT), C-Reactive Protein (CRP) and Interleukin 6 (IL-6). Multiple studies have demonstrated the high sensitivity of these biomarkers to sepsis. The proposed device will combine for the first time the analysis of cell-surface proteins and plasma proteins biomarkers from the same sample of blood. Such a device, combined with the routinely test performed in the hospitals, could significantly accelerate the diagnosis of sepsis and as consequence the clinical decision, to provide the correct treatment to the patients.