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

用于禽流感多靶点 qPCR 检测的新型通用探针

• 批准号: 7614848
• 负责人: DAVID A SHAFER
• 金额: $ 19.62万
• 依托单位: GENETAG TECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-09 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614848
• 关键词: Avian Influenza; Binding; Biological Assay; Cessation of life; Clinical; Code; Color; Communicable Diseases; Confidential Information; Consensus Sequence; DNA; DNA Sequence Rearrangement; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Discrimination; Doctor of Philosophy; Exhibits; Flu virus; Gene Mutation; Genes; Genomics; Genotype; Grant; HIV; Hemagglutinin; Hospitals; Infection prevention; Influenza; Label; Legal patent; Methods; Microarray Analysis; Molecular; Monitor; Morbidity - disease rate; Mutate; Mutation; Neuraminidase; Nucleic Acid Amplification Tests; Nucleic Acids; Oligonucleotides; Outcome; Patients; Pattern; Performance; Phase; Phenotype; Point Mutation; Population; Principal Investigator; Procedures; Process; Program Development; Real-Time Systems; Relative (related person); Research Personnel; Sampling; Seasons; Sensitivity and Specificity; Series; Signal Transduction; Site; Specificity; Speed; Surface Antigens; System; Technology; Testing; Time; Tube; Vaccines; Variant; Viral; Virus; Work; base; color detection; comparative; cost; design; flexibility; flu; improved; innovation; mortality; new technology; novel; pandemic influenza; pathogen; prevent; prototype; public health relevance; routine practice

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.

描述（由申请人提供）：突变和遗传重排的高率，特别是对于表面抗原血凝素和神经氨酸酶，会导致主要流感病毒病毒和周期性流感大流传学的年度变化，这可能威胁整个人群。因此，需要使用诊断方法来识别和保护世界人口免受这些新出现的新菌株的影响。近年来，由于此类方法在检测基因组变化的基因组变化的功能上，病毒诊断一直在迅速转移到核酸测试（NAT），这是造成抗原变化的原因，并且可以在物种和菌株之间进行精确歧视。 NAT诊断主要集中在基于PCR的方法上，尤其是实时PCR，因为这些方法提供了样本放大和速度。但是，基于PCR的方法仅限于一些靶标和实时PCR，其假阳性和假否定率很高。研究者开发了新颖的通用探针，可以轻松地以低成本应用于任何目标，并提供了用组合信号在一个管中检测多个靶标的能力。该技术还可以实现多目标的多色检测，可提供独特的信号传导模式。这些新功能可以克服假否定性和误报，从而提高了测定可靠性。该应用计划扩展并优化了针对流感的H5，H3和M1的原型通用探针的开发，并为H1和Flu。还将进行其他探针。还将进行测试，以比较针对针对相同目标的标准Taqman探针的新通用探针的性能。这些创新方法的预期结果应产生快速，敏感和低成本的通用探测系统，以实时流感检测和歧视。公共卫生相关性：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，该技术可以同时对致病物种中多个目标位点进行低成本检测。因此，该技术可以取代当前的Taqman探测器，这些探针构成昂贵，仅检测一个序列目标。多目标检测可防止虚假的阴性测试并提高可靠性。误报也可以通过实现多色检测的技术的变化来避免。探针将通过检测H5，H3，H1和MI中的3个目标位点来检测和区分流感A的主要菌株。这些诊断探针旨在用于医院和临床实验室的商业分布，用于测试和监测传染病和管理治疗。