A Binary Probe-Based DNAzyme Cascade for Rapid Detection of MRSA/MSSA

描述(由申请人提供)：用于快速检测MRSA/MSSA的基于双探针的DNAzyme级联项目概述该STTR第一阶段项目将开发一种概念验证分子测试，用于快速检测金黄色葡萄球菌(SA)，并在没有聚合酶扩增的情况下区分耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林敏感金黄色葡萄球菌(MSSA)。二元DNAzyme探针将被设计成与MRSA特异的基因或SA特异的基因互补。在靶基因存在的情况下，双探针将组装成催化活性的DNAzyme，并激活DNAzyme级联，提供视觉或比色读数。第一阶段的研究将集中于设计双链DNAzyme探针，构建DNAzyme级联，并论证MRSA/MSSA筛选的技术可行性。与MSSA感染相比，MRSA感染的治疗选择有限，并导致显著更高的发病率、死亡率和医疗成本。因此，快速、可靠地检测金黄色葡萄球菌并区分MRSA和MSSA将具有重大的临床、经济和社会影响。如果开发成功，建议的DNAzyme级联方法将显著提高目前不含PCR的分子测试的检测极限，并有可能开发成商业上可行的、独立的单管分析方法，操作时间最短，不需要复杂的仪器。 公共卫生相关性：金黄色葡萄球菌，特别是耐甲氧西林金黄色葡萄球菌，是从皮肤和软组织感染到侵袭性、危及生命的感染的主要原因 拟议的不含聚合酶链式反应的分子检测将满足对MRSA和MSSA进行快速、敏感和特异性筛查的迫切和尚未得到满足的需求，这种筛查方法简单易用、成本效益高，从而能够及时和准确地治疗MRSA/MSSA感染，并有助于降低相关的医疗成本。

DESCRIPTION (provided by applicant): A binary probe-based DNAzyme cascade for rapid detection of MRSA/MSSA Project Summary This STTR Phase I project will develop a proof-of-concept molecular test for rapid detection of Staphylococcus aureus (SA) and differentiation between methicillin-resistant SA (MRSA) and methicillin-susceptible SA (MSSA) in the absence of PCR amplification. Binary DNAzyme probes will be designed to be complementary to an MRSA-specific gene or to an SA-specific gene. In presence of a target gene, the binary probes will be assembled into a catalytically active DNAzyme and activate a DNAzyme cascade, providing a visual or colorimetric readout. The Phase I research will focus on designing the binary DNAzyme probes, constructing the DNAzyme cascade, and demonstrating the technical feasibility for MRSA/MSSA screening. Compared to MSSA infections, MRSA infections have limited treatment options and result in significantly higher morbidity, mortality, and healthcare costs. Therefore, rapid and reliable detection of SA and differentiation between MRSA and MSSA will have significant clinical, economic, and societal impacts. If successfully developed, the proposed DNAzyme cascade approach will significantly improve the detection limit of current PCR-free molecular tests and have the potential to be developed into commercially viable, self-contained single-tube assays with minimum hands-on time and no requirement for sophisticated instrument. PUBLIC HEALTH RELEVANCE: Staphylococcus aureus, particularly MRSA, is a major cause of infections ranging from skin and soft tissue infections to invasive, life-threatening infections such as bacteremia, endocarditis, etc. The proposed PCR-free molecular test would address an urgent and unmet need for rapid, sensitive, and specific screening of MRSA and MSSA in an easy-to-use, cost- effective way, thereby allowing for timely and accurately treatment of MRSA/MSSA infections and helping reduce the associated healthcare costs.