novel universal probes for multi-target qPCR detection of avian flu

描述(申请人提供)：高突变率和基因重排，特别是表面抗原血凝素和神经氨酸酶，导致主要流感病毒和周期性流感大流行每年发生变化，可能威胁整个人口。因此，需要诊断方法来识别和保护世界人口免受这些新出现的毒株的影响。近年来，病毒诊断正在迅速转向核酸检测(NAT)，因为这种方法能够检测潜在的基因组变化，这些变化是抗遗传变化的原因，并使物种和毒株之间能够精确区分。NAT诊断主要集中在基于聚合酶链式反应的方法，特别是实时聚合酶链式反应，因为这些方法提供了样本放大和速度。然而，基于聚合酶链式反应的方法仅限于少数靶点，并且实时聚合酶链式反应存在较高的假阳性和假阴性率。研究人员已经开发出新的通用探测器，这种探测器可以很容易地以低成本应用于任何目标，并提供利用组合信号在一个管中探测多个目标的能力。这项技术还可以实现多目标、多颜色检测，从而提供独特的信号模式。这些新功能可以克服假阴性和假阳性，从而提高分析的可靠性。该应用程序计划扩展和优化针对H5、H3和M1流感病毒的原型通用探针的开发，并为H1和流感B创建额外的探针。还将进行测试，以比较新的通用探针与针对相同目标的标准Taqman探针的性能。这些创新方法的预期结果应该会产生快速、灵敏和低成本的通用探针系统，用于实时检测和区分流感。公共卫生相关性：该开发计划将一个经过验证的核酸扩增平台与研究人员发明的新探针技术相结合，能够同时、低成本地检测致病物种中的多个目标位置。因此，这项技术可以取代目前昂贵的只能检测一个序列靶标的Taqman探针。多目标检测防止了假阴性测试，提高了可靠性。假阳性也可以通过启用多颜色检测的技术的变化来避免。这些探针将通过检测H5、H3、H1和MI的3个靶点来检测和区分主要的甲型流感毒株。这些诊断探头旨在用于医院和临床实验室的商业分发，用于测试和监测传染病以及管理治疗。

DESCRIPTION (provided by applicant): The high rate of mutation and genetic rearrangement, particularly for the surface antigens hemagglutin and neuraminidase, results in annuals shifts in the predominant flu viruses and periodic flu pandemics that can threaten entire populations. Diagnostic methods are therefore needed to identify and to protect the world's populations against these emerging new strains. In recent years viral diagnostics has been rapidly shifting to nucleic acid testing (NAT) because of the power of such methods for detecting underlying genomic changes that are the cause of antigenetic changes and that enable precise discrimination between species and strains. NAT diagnostics has primarily focused on PCR-based methods, particularly real-time PCR, because these methods offer sample amplification and speed. However, PCR-based methods are limited to a few targets and real-time PCR suffers from high rates of false positives and false negatives. The investigator has developed novel universal probes that can easily be applied to any target at low cost and that provide the capacity to detect multiple targets in one tube with a combined signal. This technology also enables multi-target, multi-color detection which provides distinctive signaling patterns. These new features can overcome both false negatives and false positives, thereby increasing assay reliability. This application plans to extend and optimize development of prototype universal probes for H5, H3 and M1 of flu, and to create additional probes for H1 and flu B. Tests will also be conducted to compare the performance of the new universal probes against standard Taqman probes directed to the same targets. The expected outcome of these innovative methods should produce rapid, sensitive, and low cost universal probe systems for real-time influenza detection and discrimination. PUBLIC HEALTH RELEVANCE: The development program combines a proven nucleic acid amplification platform with new probe technologies invented by the investigator that enable simultaneous, low cost detection of multiple target sites in a pathogenic species. This technology can thus supplant current Taqman probes which are costly and detect only one sequence target. Multi-target detection prevents false negative tests and improves reliability. False positives can also be avoided by a variation of the technology that enables multi-color detection. The probes will detect and discriminate the major strains of influenza A by detecting 3 target sites in H5, H3, H1, and MI. These diagnostic probes are intended for commercial distribution in hospital and clinical labs for testing and monitoring infectious disease and for managing treatment.