Chelated Lanthanide Emission Fingerprinting for Multiplexed Homogeneous NAT

用于多重均质 NAT 的螯合镧系元素发射指纹图谱

基本信息

批准号：
8454682
负责人：
Amy Droitcour
金额：
$ 16.11万
依托单位：
WAVE 80 BIOSCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2014-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Multiplexed homogeneous bioassays provide an important alternative to microarrays for multiplexed analysis of biological samples, because they avoid some of assay development challenges and can greatly reduce assay turnaround time. For simultaneously detecting multiple targets in a homogeneous assay format, target-specific fluorescent labels (in various configurations, such as directly labeling and beacons) are an effective tool. Additional methods for multiplexed homogeneous bioassays, such as the Chelated Lanthanide Emission Fingerprinting (CLEF) method proposed here, would complement fluorescence methods and potentially offer advantages for some applications, such as those where filter wheels or multiple detectors are impractical due to the size or robustness requirements of the instrument. Multiplexed point of care diagnostic devices are improved by both the rapid turnaround time of homogeneous methods and the potential size and durability improvements achieved by avoiding filter wheels in the proposed multiplexing method. The CLEF assay scheme utilizes the long luminescent lifetimes of chelated lanthanides with molecular beacon technology to provide sensitive, multiplexed detection of nucleic acids. The assay takes place entirely in solution, speeding turnaround time. The CLEF assay uses luminescence-lifetime-based multiplexing; the different target-specific beacons each have different luminescent decay profiles, determined by the chelated lanthanide conjugated to the beacon. All the beacons are excited with a single LED, and all the beacons that are hybridized to their complementary target luminesce, while the energy of those that are not hybridized is quenched by a dark quencher. All emissions are read with a single emission filter and a single detector. The CLEF method is enabled by an instrument with a novel multiplexing algorithm and high-speed optics and data acquisition hardware. The CLEF algorithm separates the luminescent decay into the components contributed by each beacon, determines which beacons are signaling and which are quenched. In this 6-month R43 Phase I, work is organized around two specific aims designed to demonstrate the feasibility of this novel homogeneous multiplexing approach - Aim 1 establishing the reporter chemicals and conjugating them to probes, and Aim 2 enhancing the high-speed instrument and extending signal processing algorithms from duplex to greater degrees of multiplexing. 1

描述（由申请人提供）：多路复用的均匀生物测定为生物样品多路复用分析提供了一个重要的替代方法，因为它们避免了一些测定的开发挑战，并且可以大大减少测定周转时间。为了以均匀测定格式同时检测多个目标，目标特异性荧光标签（以各种配置，例如直接标记和信标）是有效的工具。多路复用均质生物测定的其他方法，例如此处提出的螯合的灯笼发射指纹（CLEF）方法，它将补充荧光方法，并可能为某些应用提供优势，例如由于仪器的大小或鲁棒性要求，滤轮轮或多个检测器是不切实际的。多路复用点的诊断设备通过均匀方法的快速周转时间以及通过在建议的多路复用方法中避免过滤器轮来改善的快速周转时间以及潜在的大小和耐用性提高。 CLEF分析方案利用了用分子信标技术的螯合灯笼的长发光寿命来提供核酸的敏感，多路复用的检测。该测定完全是在解决方案中进行的，从而超越了周转时间。 CLEF分析使用基于千篇一律的多重发光。不同目标特异性的信标每个具有不同的发光衰减曲线，这是由螯合的灯笼缀合的灯笼缀合的。所有的信标都对单个LED感到兴奋，并且所有与互补的目标发光杂交的信标都会激发，而未杂交的信标则被黑淬火器淬灭。所有排放均使用单个发射过滤器和一个单个检测器读取。 CLEF方法是通过具有新颖的多路复用算法和高速光学和数据采集硬件的仪器来启用的。 CLEF算法将发光衰减分为每个信标贡献的组件，确定信标是信号传导并淬火的。在这个为期6个月的R43阶段I期间，工作围绕两个特定的目的进行组织，旨在证明这种新型均匀的多路复用方法的可行性 - 目标1建立报告基因化学品并将其偶联以探测，并将其瞄准2增强高速仪器，并扩展了从Duplex到更大程度的多重倍增度的信号处理算法。 1