CLINICAL STUDY ON INOS ELISA FOR SEPSIS AND SEPTIC SHOCK

我们最初提出并随后在一项先导性临床研究中发现，在严重脓毒症和感染性休克发生之前的全身炎症反应综合征(SIRS)或脓毒症发作期间，循环中人类诱导型一氧化氮合酶(HiNOS)水平显著增加。我们的目标是开发临床使用的免疫分析方法，在严重脓毒症和感染性休克发生之前，快速检测、定量和监测hiNOS的这种增加，从而为未来的有害事件提供早期预警信号。早期诊断将为比目前更早开始治疗提供基础。为了实现我们的目标，我们开发了一种针对hiNOS的高度特异性的夹心ELISA法，该方法基于我们的20种抗hiNOS单抗。在对10名ICU患者进行的一项初步临床研究中，我们已经使用我们开发的用于hiNOS的ELISA和其他三种确证免疫分析方法来检测和测量外周血单核细胞(PBMC)中hiNOS的水平。这项初步研究的结果表明，可以检测到PBMC中hiNOS水平的升高，并可能成为预测严重脓毒症和感染性休克发病的良好生化标志物。这项第一阶段计划的目的是将临床研究扩展至更多的病人，以评估在先导临床研究期间所作的观察是否适用于更多的病人，以评估在先导临床研究期间所作的观察是否普遍适用。一旦完成，整个项目将带来所需的新的临床诊断标记，这反过来将导致改善危重患者的医疗保健。拟议的商业应用：在美国，每年发生超过1,200,000例全身炎症反应综合征和败血症；这导致每年超过500,000例严重败血症和感染性休克，以及每年100,000-250,000例死亡。需要一种快速测试来评估患者进入和沿着脓毒症途径进展到严重的脓毒症和感染性休克。我们的免疫检测旨在满足这一临床需求，通过减少ICU停留时间来降低患者成本，并改善患者预后。

We initially proposed and then subsequently found during a pilot clinical study that a measurable increase in the circulating level of human inducible nitric oxide synthase (hiNOS) occurs during an episode of systemic inflammatory response syndrome (SIRS) or sepsis prior to the onset of severe sepsis and septic shock. Our goal is to develop immunoassays for clinical use that can rapidly detect, quantitate and monitor this increase in hiNOS prior to the onset of severe sepsis and septic shock, and thus provide an early warning signal for future deleterious events. An early diagnosis will provide a basis for an earlier initiation of therapies than is currently practiced. To achieve our goal, we have developed a highly specific sandwich ELISA for hiNOS based upon our panel of twenty anti-hiNOS monoclonal antibodies. The ELISA and three other confirmatory immunoassays that we developed for hiNOS have been used to detect and measure the level of hiNOS in peripheral blood monocytes (PBMC) during an pilot clinical study on a cohort of 10 ICU patients. The results obtained from this pilot study indicate that an increased level of hiNOS in PBMC can be measured and may be a good biochemical marker that can be used to prognosticate the onset of severe sepsis and septic shock. The aim of this phase I project is to extend the clinical study to a larger number of patients in order to assess if the observations made during the pilot clinical study to a larger number of patients in order to assess if the observations made during the pilot clinical study are generally applicable. Once completed, the overall project will bring a needed new clinical diagnostic onto the marker, and this, in turn, will lead to improved health care for critically ill patients. PROPOSED COMMERCIAL APPLICATION: Over 1,200,000 cases/year of systemic inflammatory response syndrome and sepsis occur in the USA; these lead to over 500,000 cases/year of severe sepsis and septic shock and 100,000-250,000 deaths/year. A rapid test to assess a patient's entry into and progress down the sepsis pathway to severe sepsis and septic shock is needed. Our immunoassays are designed to meet this clinical need, to lower patient cost by reducing the length of ICU stays, and to improve patient outcome.