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）：突变和基因重排的高发生率，特别是表面抗原血凝素和神经氨酸酶，导致主要流感病毒每年发生变化，并导致周期性流感大流行，可能威胁整个人群。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新菌株的侵害。近年来，病毒诊断已迅速转向核酸检测（NAT），因为这种方法用于检测潜在的基因组变化的能力，这些基因组变化是抗原变化的原因，并且能够精确区分物种和菌株。NAT诊断主要集中在基于PCR的方法，特别是实时PCR，因为这些方法提供了样品扩增和速度。然而，基于PCR的方法限于少数靶标，并且实时PCR遭受高假阳性率和假阴性率。研究人员开发了新型通用探针，可以很容易地以低成本应用于任何目标，并提供了在一个管中检测多个目标的能力。该技术还能够实现多目标、多色检测，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，从而提高检测可靠性。本申请计划扩展和优化流感病毒H5、H3和M1的原型通用探针的开发，并创建H1和流感病毒B的其他探针。还将进行测试，以比较新的通用探针与针对相同靶标的标准Taqman探针的性能。这些创新方法的预期结果应该产生用于实时流感检测和区分的快速、灵敏和低成本的通用探针系统。公共卫生关系：该开发计划将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，能够同时低成本检测致病物种中的多个靶位点。因此，该技术可以取代目前昂贵且仅检测一个序列靶标的Taqman探针。多目标检测可防止假阴性测试并提高可靠性。假阳性也可以通过使多色检测成为可能的技术的变化来避免。探针将通过检测H5、H3、H1和MI中的3个靶位点来检测和区分甲型流感的主要毒株。这些诊断探头预期用于医院和临床实验室的商业销售，用于检测和监测传染病以及管理治疗。