Rapid point-of-care detection of Trichomonas Vaginalis

快速即时检测阴道毛滴虫

• 批准号: 10699836
• 负责人: Sam Bae
• 金额: $ 27.57万
• 依托单位: ELECTRONUCLEICS, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-07 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10699836
• 关键词: Achievement; Acute; Address; Adsorption; Antibiotics; Base Sequence; Binding; Biological Assay; Blood; Blood capillaries; Cells; Charge; Chemicals; Chemistry; Chlamydia trachomatis; Clinical; Clinical Microbiology; Collaborations; Communicable Diseases; Complex; Counseling; Cytolysis; DNA; Detection; Development; Devices; Diagnostic Equipment; Diameter; Drops; Electrophoresis; Event; Glass; Goals; Gonorrhea; Human; Immobilization; Infection; Laboratories; Lateral; Legal patent; Liquid substance; Marketing; Measures; Membrane; Microfluidics; Movement; Neisseria gonorrhoeae; Nucleic Acid Amplification Tests; Nucleic Acid Binding; Nucleic Acid Hybridization; Nucleic Acids; Nucleotides; Nylons; Optics; Oral cavity; Pathogen detection; Peptide Nucleic Acids; Performance; Phase; Point-of-Care Systems; Polymerase; Polystyrenes; Process; Protocols documentation; Pump; RNA; Reaction; Reagent; Resistance; Ribosomal RNA; Sampling; Scheme; Sensitivity and Specificity; Sexually Transmitted Diseases; Signal Transduction; Specificity; Sputum; Swab; System; Technology; Testing; Time; Trichomonas Infections; Trichomonas vaginalis; Urine; Vagina; Work; analog; base; comparative; cost; cost effective; data acquisition; design; detection platform; detector; disease diagnostic; electric field; feasibility testing; inhibitor; lateral flow assay; novel; nucleic acid detection; nucleic acid purification; pathogen; point of care; point-of-care detection; point-of-care diagnostics; prototype; research and development; response; sensor; sensor technology; success

Project Summary The goal of this project is to demonstrate feasibility of a system for the point-of-care (POC) detection of Trichomonas vaginalis (TV) in human vaginal swab samples that integrates our patented detector of pathogens based on identification of species-specific, nucleic acid (NA) sequences. The detector relies on a novel electromechanical signal transduction mechanism that enables the low-cost, optics-free and amplification-free (e.g., no PCR) detection of DNA/RNA at ultralow concentration (~10-19 M). A compelling need persists for rapid (minutes), cost effective, POC NA detection devices for infectious disease diagnostics so as to facilitate prompt, well-informed therapies and counseling and to avoid potentially harmful actions including the inappropriate prescription of antibiotics. In earlier work, we demonstrated the detection of Neisseria gonorrhoeae (NG) with sensitivity of ~98% and specificity of ~100%. Trichomoniasis, the most common curable, sexually transmitted disease in the US, is often tested for in parallel with NG and Chlamydia trachomatis (CT) infections and is therefore a logical target for us to address. A key feature of the patented detector is the use of peptide nucleic acid (PNA) capture probes, which are uncharged polyamide analogs to NAs that share the same base chemistry. Since bead-PNA conjugates are designed to be charge neutral, they do not exhibit electrophoretic movement in the presence of a DC electric field. However, the substantial negative charge acquired upon capture of a target NA sequence makes the hybridized conjugate mobile. Electrophoresis of the bead-PNA conjugate with hybridized target NA to the mouth of a smaller diameter glass pore causes a significant increase in pore resistance, thereby resulting in a strong, sustained drop in measured ionic current. Nonspecifically bound NA is removed from the bead conjugate in the strong electric field at the pore mouth resulting in no sustained signal. Further, the opposing electroosmotic flow through the glass pore sweeps PNA-bead conjugates without hybridized target away from the pore mouth. In such a way, this simple conductometric device gives a highly selective (no observed false positives), binary response signaling the presence or absence of the target NA (and associated pathogen). This project focuses on two Aims: 1) Demonstration of the detection of TV in remnant vaginal swab samples from the UCLA Clinical Microbiology Laboratory with competitively high sensitivity (≥95%) and specificity (≥95%) and 2) Development and feasibility testing of key steps in an integrated and robust laboratory prototype device for rapid (<5 mins) pathogen detection based on the relatively inexpensive lateral flow assay (LFA) format. Successful achievement of these Aims will result in a device concept ready for manufacturable prototype development that is capable of NG and TV testing with competitive accuracy and likely with significant advantages of cost, rapidity, power and robustness. Further research and development work is envisioned to expand sampling capabilities to other pathogens, including CT, and to other complex sample media including blood and sputum.

项目摘要 该项目的目的是证明系统的可行性（POC）检测 人类阴道拭子样品中阴道（电视），整合了我们专利的病原体检测器 基于规格特异性的核酸（NA）序列的鉴定。探测器依靠小说 机电信号转导机制，可实现低成本，无光学和无放大 （例如，无PCR）在超高浓度（〜10-19 m）下检测DNA/RNA。迫切需要的需要 （分钟），具有成本效益的POC NA检测设备，用于传染病诊断，以促进及时， 信息良好的疗法和咨询，并避免潜在的有害行动，包括不适当的行动 抗生素的处方。在较早的工作中，我们证明了淋病奈瑟氏菌（NG）的检测 敏感性约为98％，特异性约为100％。 trichomoniasis，最常见的治愈性，性传播 在美国疾病通常与NG和衣原体沙眼（CT）感染并行测试，IS 因此，我们可以解决的逻辑目标。专利探测器的关键特征是使用胡椒核 酸（PNA）捕获问题，这些问题是共有相同基础化学的NA的无负荷的聚酰胺类似物。 由于珠-PNA结合物的设计为中性，因此不会杀死电泳运动 直流电场的存在。但是，捕获目标时获得的大量负电荷 NA序列使杂交共轭移动移动。珠-PNA的电泳与 杂交靶NA到较小直径玻璃孔的口，孔显着增加 电阻，从而导致测得的离子电流持续下降。非特定绑定的na是 从孔口的强电场中的珠子缀合物中移出，导致没有持续信号。 此外，相反的电子渗透流穿过玻璃孔糖糖pna珠偶联物 杂交目标远离孔口。通过这种方式，这个简单的导体设备可提供高度 选择性（未观察到的假阳性），二进制响应信号表明靶NA的存在或不存在 相关病原体）。该项目侧重于两个目的：1）证明电视在残留物中的检测 来自UCLA临床微生物学实验室的阴道拭子样品具有竞争性高灵敏度（≥95％） 和特异性（≥95％）和2）在集成且健壮的关键步骤的发展和可行性测试 基于相对廉价横向的快速（<5分钟）病原体检测的实验室原型装置 流量测定（LFA）格式。这些目标的成功实现将导致设备概念准备 能够具有竞争力的精确性和可能 具有成本，速度，权力和鲁棒性的显着优势。进一步的研发工作是 设想将采样能力扩展到其他病原体，包括CT和其他复杂的样本介质 包括血液和痰。