Ultrasensitive Point of Care Antigen Test for Detection of Neisseria Gonorrhoeae Using Plasmonic Flours

使用等离子面粉检测淋病奈瑟菌的超灵敏护理点抗原测试

• 批准号: 10726196
• 负责人: Sumanth Gandra
• 金额: $ 20.35万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10726196
• 关键词: Accounting; Address; Antigen Presentation; Antigens; Antimicrobial Resistance; Biological Assay; COVID-19 detection; Cells; Centers for Disease Control and Prevention (U.S.); Characteristics; Chlamydia; Clinical; Clinical Sensitivity; Communicable Diseases; Development; Devices; Diagnosis; Diagnostic; Diagnostic tests; Enzyme-Linked Immunosorbent Assay; Equipment; Feasibility Studies; Flour; Fluorescence; Future; Generations; Goals; Gonorrhea; Health; Hospitals; Human Resources; Incidence; Infection; Investigation; Label; Laboratories; Life; Lipoproteins; Methods; Microbiology; Molecular; Nature; Neisseria gonorrhoeae; Nucleic Acid Amplification Tests; Nucleocapsid Proteins; Outcome; Patients; Persons; Process; Public Health; Rapid diagnostics; Reader; Reporting; Reproducibility; Research; Resolution; Resource-limited setting; Resources; Running; SARS-CoV-2 antigen; Sampling; Sensitivity and Specificity; Sexually Transmitted Diseases; Signal Transduction; Specificity; Specimen; Speed; Surface; Technology; Testing; Time; Trichomonas Infections; United States; Urine; Validation; Work; antigen detection; antigen test; cost; detection assay; detection limit; detection test; diagnostic accuracy; diagnostic technologies; fluorophore; follower of religion Jewish; healthy volunteer; improved; infection rate; innovation; instrumentation; lateral flow assay; low and middle-income countries; nano; nanoGold; nanolabel; novel; plasmonics; point of care; point of care testing; portability; prototype; repository; screening; skills; test strip; tool

Project Summary/Abstract: An antigen test for Neisseria gonorrhoeae (Ng) that is accurate, simple, scalable, non-invasive, rapid, and effective at the point-of-care (POC), would transform public health decisions. Although antigen detection rapid diagnostic tests (Ag-RDT's) for Ng are available, they have poor sensitivity (50% or less) compared to gold standard nucleic acid amplification tests (NAAT). A NAAT based POC test was recently cleared by the US FDA, however its use in resource-poor settings is limited by its high cost. To overcome these barriers for the first time, we will harness ultrabright fluorescent nanoconstructs, called plasmonic-fluors, a recent breakthrough from our labs that enables such antigen testing with sensitivity that matches NAATs. The objective of this proposal is to develop an ultrasensitive plasmon-enhanced lateral flow assay (p-LFA) that detects the lipoprotein H.8 antigen present on Ng surface. The rationale underlying this proposal is that a highly sensitive Ag-RDT that can match the accuracy of NAAT could meet the critical need for diagnosing Ng infection at the POC and in resource-limited settings. We will achieve this by pursuing two specific aims: 1) Develop and optimize the p-LFA for detecting Ng lipoprotein H.8 antigen in urine samples obtained from healthy people. 2) Determine the diagnostic accuracy of the Ng antigen test compared to gold standard NAAT in urine specimens from patients with suspected Ng infection. The proposed approach is innovative in that it harnesses plasmonic-fluor as a fluorescent nanolabel in a LFA as opposed to the conventional gold nanoparticles, which provide only a weak colorimetric signal. With over 1000-fold improvement in sensitivity, this approach has the potential to transform antigen testing. The proposed research is significant as the novel p-LFA will meet the critical needs for widespread Ng testing scalability, speed, and low cost. The expected outcome of this work is a simple, non-invasive, Ag- RDT for Ng infection, implemented as a LFA that can replace NAAT by virtue of its accuracy and low cost in resource-limited settings. This test will have a tremendous positive impact immediately as it will be applicable to large scale Ng testing with an inexpensive test strip and minimal battery-powered portable equipment, without relying on skilled personnel. Furthermore, this test will serve as a prototype for ultrasensitive antigen tests applicable to many other infectious diseases in addition to the possibility of incorporating multiple antigens on a LFA for diagnosing other sexually transmitted infections such as chlamydia and trichomoniasis in addition to Ng.

项目摘要/摘要： 淋病奈瑟菌 (Ng) 抗原检测准确、简单、可扩展、非侵入性、快速且可靠 在护理点（POC）有效，将改变公共卫生决策。虽然抗原检测速度很快 Ng 的诊断测试 (Ag-RDT) 可用，但与金相比，它们的灵敏度较差（50% 或更低） 标准核酸扩增测试（NAAT）。基于 NAAT 的 POC 测试最近获得美国 FDA 批准， 然而，其在资源贫乏地区的使用因其高成本而受到限制。为了第一次克服这些障碍， 我们将利用超亮荧光纳米结构，称为等离子体荧光，这是我们最近的突破 实验室能够以与 NAAT 相匹配的灵敏度进行此类抗原检测。该提案的目的是 开发可检测脂蛋白 H.8 抗原的超灵敏等离子增强侧流测定 (p-LFA) 存在于 Ng 表面。该提案的基本原理是高度敏感的 Ag-RDT 可以匹配 NAAT 的准确性可以满足在 POC 和资源有限的情况下诊断 Ng 感染的关键需求 设置。我们将通过追求两个具体目标来实现这一目标：1）开发和优化用于检测的 p-LFA 健康人尿液样本中的 Ng 脂蛋白 H.8 抗原。 2）确定诊断准确性 疑似 Ng 患者尿样中 Ng 抗原检测与金标准 NAAT 的比较 感染。所提出的方法是创新的，因为它利用等离子体荧光作为荧光纳米标记 LFA 与传统的金纳米粒子不同，后者仅提供微弱的比色信号。和 这种方法的灵敏度提高了 1000 倍以上，有可能改变抗原检测。这 拟议的研究意义重大，因为新型 p-LFA 将满足广泛的 Ng 测试的关键需求 可扩展性、速度和低成本。这项工作的预期结果是针对 Ng 的简单、非侵入性 Ag-RDT 感染，作为 LFA 实现，可以凭借其准确性和在资源有限的情况下的低成本来取代 NAAT 设置。该测试将立即产生巨大的积极影响，因为它将适用于大规模 Ng 使用廉价的测试条和最少的电池供电便携式设备进行测试，无需依赖熟练的技术人员 人员。此外，该测试将作为适用于许多疾病的超灵敏抗原测试的原型。 除了在 LFA 上整合多种抗原进行诊断的可能性之外，还可以用于其他传染病 除 Ng 外，其他性传播感染如衣原体和滴虫病。