Rapid Molecular Detection of PML/RARalpha

我们提出了一系列实验，旨在利用基于MEMS的电化学传感器阵列来快速检测哺乳动物细胞中两种最常见的PML-RARpha融合mRNAs。这项研究的最终目标是开发一种廉价、用户友好的系统，能够在不到一小时的时间内诊断出90%的APL病例。这样的系统将使ATRA在最初出现的几小时内启动，而不是几天，从而降低危及生命的出血风险。一旦该系统建成，这一集成系统的最大好处将是更高的完全缓解率，更重要的是，由于早期诊断，提高了APL患者的总体存活率。此外，这项新技术有望在肿瘤学领域具有越来越大的潜在用途，在肿瘤学领域，驱动恶性疾病过程的分子变化正在继续被表征。也许与人类恶性肿瘤相关的研究最深入的分子变化是在95%的慢性粒细胞白血病(CML)患者的白血病细胞中发现的Philadelphia染色体，这导致了bcr和abl基因的融合。我们在这里开发的检测系统有望成为CML和其他恶性肿瘤的快速诊断系统。癌基因或抑癌基因表达改变、易位和其他易感基因改变的分子诊断将越来越多地用于癌症检测和治疗方案的选择。基于MEMS-0的手持式电化学阵列检测的速度、通用性、便携性、易用性和成本是当前分子技术无法比拟的显著优势。原型PML/RARpha传感器阵列的分析有效性将通过第一阶段的里程碑得到验证。建议的商业应用：GeneFluidics的生命科学/电化学实验室位于加利福尼亚州洛杉矶的商业/工业区--加利福尼亚州托兰斯。这个地点距离加州大学洛杉矶分校有20分钟的路程，欧文·陈博士和尼尔·沙阿博士的办公室就在那里。我们的实验室占地1000平方英尺。研发用台式电脑4台，笔记本电脑3台，具备完整的互联网接入和网络连接能力。我们实验室有一个机械加工和电子样机部门。隔离室是为“肮脏”的加工而建造的，并且房间是加压的，这样它就不会污染隔离室外的实验装置。

We propose a series of experiments aimed to develop rapid detection of two most common PML-RARalpha fusion mRNAs from mammalian cells using MEMS-based electrochemical sensor array. The ultimate goal of the studies is the development of an inexpensive, user-friendly system capable of diagnosing 90 percent of APL cases in less than one hour. Such a system would facilitate initiation of ATRA within hours, instead of days, of initial presentation, and thereby reduce the risk of life- threatening hemorrhage. Once the system is completed the greatest benefits of this integrated system will be higher complete remission rates, and more importantly, improved overall survival of patients with APL due to earlier diagnosis. Furthermore, this novel technology promises to be of growing potential use to the field of oncology, where the molecular alterations which drive malignant diseases processes are continuing to be characterized. Perhaps the most well-studied molecular alteration associated with human malignancy is the Philadelphia chromosome found in the leukemic cells of >95% of patients with chronic myelogenous leukemia (CML), which results in the fusion of BCR and ABL genes. The detection system we aim to develop here is expected to be made amenable to the rapid diagnosis of CML and perhaps other malignancies as well. Molecular diagnosis of alterations in oncogene or tumor suppressor gene expression, translocations, and others predisposing genetic alterations will increasingly be used for cancer detection and choice of therapeutic options. The speed, versatility, portability, ease of use and cost of the handheld MEMS- 0based electrochemical array detection are significant advantages over current molecular technologies. The analytical validity of a prototype PML/RARalpha sensor array will be shown through milestones in Phase I. PROPOSED COMMERCIAL APPLICATIONS: GeneFluidics' Life Science/Electrochemistry Laboratory is located in Torrance, CA, a business/industrial area of Los Angeles, CA. This location is 20 minutes from UCLA where Dr. Irvin Chen and Dr. Neil Shah have their office. Our laboratory occupies 1,000 square feet. For research and development purposes, there are 4 desktop computers, 3 laptop computers and complete internet access and network connection. Our laboratory has a section for machining and electronics prototyping. An isolation room is built for "dirty" machining and the room is pressurized so it won't contaminate the experiment setup outside the isolation room.