Low-Cost Instrument-free Point-of-Care Diagnostic for Neisseria gonorrhoeae

低成本、免仪器的淋病奈瑟氏菌即时诊断

• 批准号: 9256272
• 负责人: DEBORAH Anne DEAN
• 金额: $ 71.67万
• 依托单位: LUCIRA HEALTH, INC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-25 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9256272
• 关键词: Accident and Emergency department; Antibiotics; Bacterial Sexually Transmitted Diseases; Bedside Testings; Binding Sites; Biological Assay; Blood; Care Technology Points; Centers for Disease Control and Prevention (U.S.); Cervical; Chemistry; Chlamydia trachomatis; Chronic; Cities; Clinic; Clinical; Clinical Trials; Clinics and Hospitals; Computer Simulation; Cytolysis; DNA; Data; Databases; Detection; Development; Diagnosis; Diagnostic; Drug resistance; Early Diagnosis; Ectopic Pregnancy; Ensure; Epidemic; Equipment; Family; Genome; Genomics; Goals; HIV-1; Head; Hospitals; Human; Infection; Infertility; Lead; Leukocytes; Medical; Microfluidics; Modification; Mucous body substance; Multi-Drug Resistance; Multi-Institutional Clinical Trial; Neisseria gonorrhoeae; Nucleic Acid Amplification Tests; Nucleic Acids; Organism; Pathway interactions; Patients; Pelvic Inflammatory Disease; Pelvis; Phase; Physicians; Point-of-Care Systems; Predictive Value; Preparation; Prevalence; Privatization; RNA; Recovery; Research; Resources; Rural; Sampling; Seminal fluid; Sensitivity and Specificity; Sexually Transmitted Diseases; Site; Specimen; Swab; System; Technical Expertise; Teenagers; Temperature; Testing; TimeLine; Training; Tube; Urethra; Vagina; base; chronic pelvic pain; cost; cross reactivity; design; diagnostic screening; follow-up; geographic population; improved; inner city; innovation; instrument; instrumentation; laboratory equipment; multiplex detection; pathogen; point of care; point-of-care diagnostics; prototype; rapid detection; resistant strain; screening; stem; transmission process; user-friendly

Abstract Neisseria gonorrhoeae (GC) is the second most common cause of bacterial sexually transmitted diseases (STD) with an estimated world prevalence of 78 million. Approximately 820,000 cases occur annually in the U.S. but over half are unreported, and rates continue to rise as indicated by a 10.5% increase from 2010 to 2014. GC is a major cause of pelvic inflammatory disease (PID) that can lead to tubal factor infertility, ectopic pregnancy and chronic pelvic pain. GC also facilitates the transmission of HIV-1 infection. While we rely on antibiotics to treat GC infections, empiric therapy has fueled the already problematic issue of drug-resistant GC in the U.S. and the world. The main obstacle to stemming the spread of GC infections is the lack of a point-of- care (POC) diagnostic that could provide surveillance and early detection, reducing infection rates and sequelae. Current GC diagnostics rely on commercial nucleic acid amplification tests (NAATs) that are expensive, require equipment and highly trained operators, take a day to days for results, and can result in loss to follow up of patients. Thus, current NAATs are not suitable at the POC. Our team comprised of Drs. Dean and Gaydos, experts on STDs and POC development, and Diassess, a startup company with proprietary technology for POC diagnostics, have preliminary data showing that we: 1) can rapidly extract GC nucleic acids from endocervical swabs with no instruments or trained operator in a prototype Sample Preparation Module; 2) have an instrument-free multiplexed Detection Module for colorimetric detection of GC nucleic acids; 3) have a limit of detection of 10 organisms/assay; 4) have validated assays to detect GC strains and assays to detect human RNA/DNA with no cross reactivity with other STD pathogens; and 5) have shown that our system can detect GC reliably in clinical endocervical swabs that are know positive by a commercial NAAT. We will expand on our preliminary studies with the following aims. Aim 1: Using the expanding aggregate of reference and clinical GC genome sequences, we will refine our assays, replacing failed assays as needed, and ensure that our assays detect diverse GC clinical strains without cross-reactivity with STD pathogens or common vaginal/cervical species; Aim 2: Optimize sample preparation chemistry for nucleic acid extraction from urethral, vaginal and endocervical swabs, assay design and colorimetric chemistry for these swab types, without false negative or positive results from interfering substances; Aim 3: We will evaluate the sensitivity and specificity, and positive and negative predictive values of our fully-integrated system (the combined Sample Preparation Module with Detection Module) compared to commercial NAATs. By the end of Phase II, we will be poised to manufacture and use our rapid (<35 min), inexpensive, user-friendly, instrument-free, sensitive and specific GC POC test for clinical trials in the U.S. to obtain FDA regulatory clearance. Our overall goal is to deploy our GC POC diagnostic for use in doctor’s offices, small to large city and rural clinics, teen, family and STD clinics, ERs and hospital clinics, other testing sites, and resource-constrained settings around the world.

抽象的 淋病奈瑟菌 (GC) 是细菌性传播疾病的第二常见原因 据估计，全世界每年约有 820,000 例 STD 病例。 美国，但超过一半未报告，且比率持续上升，从 2010 年到 2010 年增加了 10.5% 2014. GC 是盆腔炎 (PID) 的主要原因，可导致输卵管因素不孕、异位 怀孕和慢性盆腔疼痛也会促进 HIV-1 感染的传播。 抗生素治疗GC感染，经验性治疗加剧了耐药GC这一本已成问题的问题 在美国和世界范围内，阻止GC感染传播的主要障碍是缺乏控制点。 护理（POC）诊断，可以提供监测和早期发现，降低感染率和 目前的 GC 诊断依赖于商业核酸扩增测试 (NAAT)。 昂贵，需要设备和训练有素的操作员，需要一天到几天的时间才能得到结果，并可能导致损失 因此，目前的 NAAT 不适合由 Dean 博士组成的团队。 和 Gaydos，STD 和 POC 开发方面的专家，以及 Diassess，一家拥有专有技术的初创公司 用于POC诊断的技术，初步数据表明我们：1）可以快速提取GC核酸 在原型样品制备中无需任何仪器或训练有素的操作员即可从宫颈内拭子中提取酸 模块；2) 具有用于GC核酸比色检测的免仪器多重检测模块 酸；3) 检测限为 10 种生物体/检测；4) 具有检测 GC 菌株的经过验证的检测方法； 检测人类 RNA/DNA 且与其他 STD 病原体无交叉反应的检测结果表明； 我们的系统可以可靠地检测商业 NAAT 检测呈阳性的临床宫颈内拭子中的 GC。 我们将扩大我们的初步研究，目标如下：利用不断扩大的总量。 参考和临床 GC 基因组序列，我们将改进我们的检测，根据需要替换失败的检测， 并确保我们的检测方法能够检测多种 GC 临床菌株，且不会与 STD 病原体发生交叉反应或 常见阴道/宫颈物种；目标 2：优化核酸提取的样品制备化学方法 来自尿道、阴道和宫颈内拭子、这些拭子类型的检测设计和比色化学， 没有干扰物质造成的假阴性或阳性结果；目标 3：我们将评估灵敏度； 我们完全集成的系统的特异性以及阳性和阴性预测值（组合 与商业 NAAT 相比，阶段结束时的样品制备模块和检测模块。 II，我们将准备制造和使用我们的快速（<35 分钟）、廉价、用户友好、无需仪器、 敏感且特异的 GC POC 测试在美国进行临床试验，获得 FDA 的整体监管许可。 目标是部署我们的 GC POC 诊断，用于医生办公室、小到大城市和农村诊所、青少年、 家庭和性病诊所、急诊室和医院诊所、其他检测场所以及周围资源有限的环境 世界。