An in vitro diagnostic assay for the early and accurate detection of platelet-activating antibodies associated with Heparin-induced Thrombocytopenia

项目摘要/摘要 该二期SBIR方案旨在开发一种高精度的功能性体外诊断方法 肝素诱导的血小板减少症(HIT)病原性抗体的IVD检测 肝素治疗的不良反应。这项检测是基于最近的发现，即血小板因子4 (PF4)处理的血小板可用于敏感和特异地检测临床上有意义的命中 抗体。HIT在美国每天导致5名以上患者死亡，并经常被怀疑使用肝素治疗 可能有多种潜在原因导致血小板减少的住院患者。协助，协助 在诊断和管理方面，医生依赖于两个HIT分析家族。第一种是PF4/聚阴离子ELISA- 基于体外诊断(IVD)的分析是敏感的，但高度非特异性，因此阳性预测 这些分析的价值很低，只有30-50%。第二，更准确的基于血小板激活的(功能) 5-羟色胺释放试验(SRA)等分析技术复杂，仅在少数情况下进行 参考实验室，这可能会导致较长的周转时间。因此，前线ELISA被用来 处理大多数疑似HIT患者和许多ELISA假阳性患者得到不适当的治疗 使用替代抗凝剂，这些抗凝剂价格昂贵，出血情况明显更糟。雷瑟姆氏病 目标是通过取代不准确的ELISA和技术复杂的SRA来革命性地进行HIT诊断 使用HITDx，一种简单但准确的功能性IVD，可以在医院实验室进行。HITDx是 基于突破性研究表明致病的血小板激活HIT抗体可以结合 并以非肝素依赖的方式激活经PF4处理的血小板。临床研究包括一名409名患者， 前瞻性、多中心、盲目研究表明，这项技术可以被利用来提供准确的 命中诊断。在第一阶段，Retham Technologies证明了长期(12个月)治疗的PF4 稳定的血小板可以偶联到基于ELISA的终点，用于检测新的分析物，展示 这项技术可以应用于IVD检测。涵盖这项技术的基础专利已经 目前已发布多项专利，在不同司法管辖区还有多项新的专利正在申请中。在这份SBIR第二阶段提案中， Retham将开发、验证和评估使用PF4的独立HIT IVD原型的性能- 治疗长期储存的血小板。HITDx在疫苗诱导抗体检测中的应用 血栓性血小板减少症是新冠肺炎疫苗的一种新并发症，将在#年进行评估 未来的研究。该项目将由柯蒂斯·琼斯领导，他是这项技术的发明者，也是 开发稳定血小板的新方法。他将得到领衔热门人物帕德马纳班博士的支持 专家、QA/RA和生物统计顾问，以及专家Retham顾问委员会成员。预计 这款影响患者的产品将通过分散功能测试和提供 快速、准确的结果将有助于早期和适当的患者管理。

PROJECT SUMMARY/ABSTRACT This phase II SBIR proposal is aimed at developing a highly accurate functional in vitro diagnostic (IVD) assay for the detection of pathogenic antibodies in Heparin-induced thrombocytopenia (HIT), an adverse reaction to heparin treatment. This assay is based on recent findings that Platelet Factor 4 (PF4)-treated platelets can be used for the sensitive and specific detection of clinically-significant HIT antibodies. HIT kills more than 5 patients every day in the US and is frequently suspected in heparin-treated hospitalized patients who may have a number of potential causes for thrombocytopenia. To assist with diagnosis and management, physicians rely on two families of HIT assays. The first, the PF4/Polyanion ELISA- based in vitro diagnostic (IVD) assays are sensitive but are highly non-specific such that the positive predictive value of these assays is poor at only 30-50%. The second, more accurate platelet activation-based (functional) assays such as the Serotonin release assay (SRA) are technically complex and are performed only at a few reference laboratories which can lead to a long turnaround times. As a result, frontline ELISAs are used to manage most HIT-suspected patients and many patients with false-positive ELISAs are inappropriately treated with alternative anticoagulants which are expensive and have a significantly worse bleeding profile. Retham’s goal is to revolutionize HIT diagnosis by replacing both the inaccurate ELISA and technically complex SRA with HITDx, a simple yet accurate functional IVD that can be performed in the hospital laboratory. HITDx is based on groundbreaking research that suggests that pathogenic platelet-activating HIT antibodies can bind and activate PF4-treated platelets in a heparin-independent manner. Clinical studies including a 409-patient, prospective, multicenter, blinded study demonstrate that this technology can be leveraged to provide accurate HIT diagnosis. During phase I, Retham Technologies demonstrated that PF4 treated, long term (12-months) stabilized platelets can be coupled to an ELISA-based endpoint for detection of a novel analyte, demonstrating that this technology can be adapted to an IVD assay. The foundational patents covering this technology have issued, and multiple additional patents are pending in various jurisdictions. In this SBIR Phase II proposal, Retham will develop, verify and assess the performance of a self-contained HIT IVD prototype using PF4- treated long-term stored platelets. The utility of HITDx for detection of antibodies that cause vaccine-induced thrombotic thrombocytopenia, a newly recognized complication of COVID-19 vaccines will be assessed in future studies. The project will be led by Curtis Jones who is inventor of this technology and spearheaded the development of novel methods to stabilize platelets. He will be supported by Dr. Padmanabhan, a leading HIT expert, QA/RA and biostatistics consultants, and expert Retham advisory board members. It is expected that this patient-impacting product will revolutionize HIT diagnosis by decentralizing functional testing and providing rapid, accurate results that will facilitate early and appropriate patient management.