Rapid No-Spin Chlamydia RNA Extraction via Filtering, Lysis and Monolith Binding

通过过滤、裂解和整体结合快速无旋转衣原体 RNA 提取

• 批准号: 8715634
• 负责人: Amy Droitcour
• 金额: $ 30万
• 依托单位: WAVE 80 BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-15 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8715634
• 关键词: Address; Adoption; Antibiotics; Antigens; Bacteria; Bacterial DNA; Bacterial RNA; Binding; Biological Assay; Blindness; Blood capillaries; Caring; Centrifugation; Chemistry; Chlamydia; Chlamydia trachomatis; Clinic; Clinical; Clinics and Hospitals; Communicable Diseases; Conjunctivitis; Cytolysis; DNA; Detection; Development; Diagnosis; Engineering; Environment; Exhibits; Female; Filtration; Flushing; Glass; HIV; Individual; Infant; Infection; Infertility; Laboratories; Lead; Life; Methods; Mothers; Nucleic Acid Amplification Tests; Nucleic Acids; Outcome; Patients; Pelvic Inflammatory Disease; Performance; Phase; Polymers; Porosity; Positioning Attribute; Pregnant Women; Preparation; Procedures; Process; RNA; Reagent; Recovery; Reporting; Ribosomal RNA; Running; Sampling; Sensitivity and Specificity; Site; Solid; Staging; Swab; System; Technology; Test Result; Testing; Time; Urine; Vagina; Validation; Viral Load result; Visit; Woman; Work; capillary; clinical decision-making; cost; design; follow-up; high risk; improved; innovation; innovative technologies; instrument; manufacturing process; meter; programs; public health relevance; sample collection; screening; usability

DESCRIPTION (provided by applicant): This application addresses the urgent need for expanded screening and test-and-treat programs for Chlamydia trachomatis (CT) through the development of a fast, straightforward sample preparation method. CT infection, which is often asymptomatic in female patients, can result in pelvic inflammatory disease and irreversible infertility in the infected individual as well as blindness in infants born to infected mothers. CT infection can be readily resolved through a short course of antibiotics, but achieving optimal patient outcomes depends on identifying infection and prescribing treatment within a single visit to a doctor's office, hospital, or clinic-the "test-and-treat" approach to infectious disease care. Multiple studies have shown current rapid CT antigen tests to exhibit such poor sensitivity as to have almost no value in clinical decision making. In contrast, both the sensitivity and the specificity of nucleic acid testing for CT infection typically exceed 95%. With increasing availability and low cost of compact thermocyclers and other emerging nucleic acid testing technologies, smaller clinics are increasingly well positioned to run the amplification and detection steps in CT nucleic acid testing. Yet the requisite preparation of vaginal swab and urine samples for subsequent CT nucleic acid testing-including lysis, filtering, and extraction and elution of bacterial RNA and DNA-is a barrier to widespread adoption of CT nucleic acid testing outside of sophisticated laboratories. This application seeks to enable expanded access to CT nucleic acid testing through the development of high- performance, fully automated CT sample preparation in an inexpensive, disposable cartridge. The usage procedure consists of simply placing a urine or vaginal swab sample in a small cup, which attaches to the cartridge. Slit capillary array fluidic actuator (SCAFA) chips-an innovative technology recently developed by the Applicant Organization-are built into the disposable cartridge and drive all sample and reagent transport steps entailed in a high-extraction-efficiency sample preparation process. The extraction process will leverage an innovative, fully cartridge-integrated (no centrifugation) soli phase extraction method using porous polymer monoliths within short glass capillaries, tuned to have optimal porosity for efficient capture of a nucleic acid target without clogging. The sample preparation process executed within the cartridge also includes pre- extraction steps of sample metering, filtering CT from sample, lysing CT on the filter, and elution from the filter. Sample-in to results-out turnaround time is 45 minutes. This Phase I project is organized around two specific aims. Specific Aim 1 focuses on the sample prep module's front end, including metering and lysing. Specific Aim 2 focuses on high-yield extraction in the porous polymer monolith, including monolith composition and capture/elution chemistry. The two Aims collectively establish the feasibility of the high-performance, low-cost CT sample preparation cartridge, laying groundwork for full development in Phase II.

描述（由申请人提供）：此申请通过开发快速，直接直接的样本准备方法来解决迫切需要扩大沙眼衣原体的筛查和测试和治疗程序。在女性患者中通常无症状的CT感染会导致受感染母亲所生的婴儿和婴儿的骨盆炎症性疾病和不可逆性不育症。 CT 感染可以通过短暂的抗生素很容易解决，但是实现最佳患者结局取决于在一次访问医生办公室，医院或临床医院或诊所中识别感染和处方治疗关心。 多项研究表明，当前的快速CT抗原测试表现出较差的敏感性，以至于在临床决策中几乎没有价值。相反，核酸测试对CT感染的敏感性和特异性通常超过95％。随着紧凑型热循环器和其他新兴核酸测试技术的可用性和低成本，较小的诊所的位置越来越好，可以在CT核酸测试中运行扩增和检测步骤。然而，对于随后的CT核酸测试，包括裂解，滤波和提取的必要制剂，用于随后的CT核酸测试和 细菌RNA和DNA的洗脱是在复杂实验室之外广泛采用CT核酸测试的障碍。该应用程序旨在通过在廉价的一次性弹药筒中开发高性能，全自动的CT样品制备来扩大对CT核酸测试的访问。使用过程包括简单地将尿液或阴道拭子样品放在一个附着在墨盒上的小杯中。 SLIT毛细管阵列液体执行器（SCAFA）芯片 - 申请人组织中的一种创新技术，该技术已内置在一次性墨盒中，并驱动所有样品和试剂运输步骤，并在高效率效率样本准备过程中需要进行。提取过程将利用短玻璃毛细管中的多孔聚合物整体堆积的创新的，完全弹药的集成型（无离心）溶作型相萃取方法，该方法具有最佳的孔隙率，可有效捕获核酸靶标，而无需堵塞。墨盒中执行的样品制备过程还包括样品计量的预萃取步骤，从样品中过滤CT，滤波器上的CT和过滤器洗脱。样品中 结果淘汰时间为45分钟。该阶段I项目围绕两个具体目标组织。特定的目标1专注于样品准备模块的前端，包括计量和雷。特定的目标2着重于多孔聚合物整体中的高收益提取，包括整体组成和捕获/洗脱化学。这两个目的共同确定了高性能，低成本的CT样品制备弹药筒的可行性，为II阶段的全面发展奠定了基础。