Optical Encoding Technology for Viral Screening Panels

描述（由申请人提供）：在过去的十年中，平行筛选技术的发展取得了实质性的进步，这些技术旨在检测多种类型的生物威胁。一种筛选技术侧重于减轻病毒生物威胁剂造成的威胁，方法是使用多重PCR，然后在Luminex的光学编码珠上进行杂交。然而，相当少的可用的头光学码（~30-40）和精密的流式细胞仪为基础的检测器，防止检测所有的病毒威胁在一个面板或使用屏幕以外的受控实验室环境。为了解决这些问题，一个基于Parallume （www.parallume.com）光学编码的系统，可以支持数千个光学编码，将被用于对珠子进行编码，这些珠子将被一个完全便携式的电池供电的读取器读取。最终目标将是开发一种筛选小组，能够使用便携式检测系统在任何地点检测、区分和识别所有已知的病毒家族。

DESCRIPTION (provided by applicant): The last decade has witnessed substantial improvement in the development of parallel screening technologies designed to detect many types of biological threats. One type of screening technology has concentrated on mitigating threats caused by viral Biothreat agents by using multiplex PCR followed by hybridization onto optically encoded beads from Luminex. However, the rather small number of usable bead optical codes (~30-40), and the delicate flow cytometer based detector, prevent the detection of all viral threats in one panel or use of the screen outside of a controlled laboratory environment. To address these issues, a system based on Parallume (www.parallume.com) optical encoding, which can support thousands of optical codes, will be used to encode beads that will be read by a completely portable, battery-powered reader. The final goal will be to develop a screening panel capable of detecting, differentiating and identifying all known viral families in any location by using a portable detection system. In collaboration with scientists at Lawrence Livermore National Laboratory (LLNL), Parallel Synthesis Technologies, Inc. (PSTI) will formulate a Parallume-encoded bead set containing 15 optical codes which will be used in a direct comparison against a 15-bead Luminex-based Viral Screening Panel (VSP) already in use at LLNL to detect human influenza viruses. Using biotinylated primers designed by the LLNL Bioinformatics Group, PCR for 15 regions of viral genomes are performed in a single reaction. A target region of each PCR product contains a sequence which is hybridized to a capture probe on the optically encoded bead. By treating the PCR product after hybridization with a streptavidin-phycoerythrin (SAPE) conjugate to label the biotinylated primers, the presence of the viral genome is detected by a serially diluted, threshold SAPE level on an encoded bead as compared to the controls. In the Phase I effort, Parallel and LLNL will compare the state-of-the-art Luminex system with the Parallume encoding technology by characterizing the same reactions on a divided PCR product mixture. The Parallel samples will be characterized using Parallume beads and Parallel's robust Multiplex Assay Reader System (MARS), which has replaced the fragile flow cytometer and lasers with micromachined silicon bead localization slides and super-bright signage LEDs, and the measurement rate, accuracy, precision and cost of the two systems directly compared. The combination of the Parallel and LLNL technologies could allow all known viruses to be screened anywhere in one PCR reaction. PUBLIC HEALTH RELEVANCE This innovation seeks to create a viral screening panel (VSP) whereby a human, animal, or plant sample containing DNA is subjected to a simultaneous screen for multiple pathogens (viruses in the current application) within a single vial. Limited virus detection technologies exist, but are hampered by technical limitations not possessed by the proposed VSP. This novel VSP proposes a method for expanding the number of viruses currently detectable in a single screen and adds to the potential functionality of the panel by being able to incorporate other screens (e.g., bacterial and other non-viral pathogens) into this single, multiplexed panel. This technology also offers a dramatically less-expensive reader (VSP analysis equipment) to accompany this new product.