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： 淋病奈瑟氏菌（NG）的抗原测试，它是准确，简单，可扩展，无创，快速和 在护理点（POC）有效，将改变公共卫生的决定。虽然抗原检测快速 可用于NG的诊断测试（AG-RDT），与黄金相比，它们的敏感性较差（50％或更低） 标准核酸扩增测试（NAAT）。美国FDA最近清除了基于NAAT的POC测试， 但是，它在资源贫乏的设置中的使用受到高成本的限制。为了第一次克服这些障碍 我们将利用Ultrabright荧光纳米构造，称为等离子 - 流体，这是最近与我们的突破 实验可以使这种抗原测试具有与NAAT相匹配的灵敏度。该提议的目的是 开发超敏感的等离子体增强的侧向流量测定法（P-LFA）检测脂蛋白H.8抗原 存在于NG表面。该提议的基本原理是可以匹配的高度敏感的Ag-RDT NAAT的准确性可以满足在POC和资源限制下诊断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）开发和优化检测P-LFA 从健康人获得的尿液样品中的Ng脂蛋白H.8抗原。 2）确定诊断精度 与怀疑NG患者的尿液标本中的金标准NAAT相比，NG抗原试验的NG抗原试验 感染。提出的方法是创新的，因为它可以利用等离氟作为荧光纳米贝尔 LFA与常规金纳米颗粒相反，该颗粒仅提供弱的比色信号。和 灵敏度提高了1000倍以上，这种方法具有转化抗原测试的潜力。这 拟议的研究很重要，因为新型P-LFA将满足广泛的NG测试的关键需求 可伸缩性，速度和低成本。这项工作的预期结果是NG的简单，无创的Ag-rdt 感染，作为LFA实施，可以通过其准确性和资源有限的低成本来代替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小电池供电的便携式设备进行测试，而无需依赖熟练的 人员。此外，该测试将用作适用于许多的超敏抗原测试的原型 除可能在LFA上掺入多种抗原以诊断的其他传染病外 除NG外，其他性传播感染（例如衣原体和毛瘤病）。