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 Neisseria Gonorrhoeae（GC）是细菌性传播疾病的第二大原因 （STD）估计的世界流行率为7800万。每年大约发生820,000例 美国，但一半以上没有报道，而且比率继续上涨，这是从2010年增长到10.5％ 2014。GC是骨盆炎症性疾病（PID）的主要原因，可能导致输卵管不育症，生态 怀孕和慢性骨盆疼痛。 GC还促进了HIV-1感染的传播。当我们依靠 治疗GC感染的抗生素，经验疗法已引发了已经有问题的耐药性GC问题 在美国和世界。阻止GC感染传播的主要障碍是缺乏 可以提供监视和早期检测的护理（POC）诊断，降低感染率和 后遗症。当前的GC诊断依赖于商业核酸扩增测试（NAATS） 昂贵，需要设备和训练有素的操作员，花一天的时间取得结果，并可能导致损失 跟进患者。那时，当前的NAAT不适合在POC上。我们的团队包括Drs。院长 Gaydos，性病和POC开发专家以及专有的初创公司DiaSsess POC诊断技术的技术具有初步数据，表明我们：1）可以快速提取GC核 在原型样品制备中，没有任何乐器或训练有素的操作员的宫颈拭子的酸 模块; 2）具有无仪器的多路复用检测模块，用于比色检测GC核。 酸； 3）对10种生物/测定的检测极限； 4）已有验证的测定法以检测GC菌株和 检测人类RNA/DNA的测定，没有与其他性病病原体的交叉反应性； 5）表明 我们的系统可以可靠地检测到临床内宫颈拭子中的GC，这些拭子被商业NAAT呈阳性。 我们将以以下目标扩展我们的初步研究。目标1：使用扩展的骨料 参考和临床GC基因组序列，我们将完善我们的评估，根据需要取代失败的测定法 并确保我们的测定法检测潜水GC临床菌株，而没有与性病病原体或 常见的阴道/宫颈物种； AIM 2：优化样品制备化学用于提取核酸 这些拭子类型的尿道，阴道和宫颈拭子，分析设计和比色化学物质， 没有干扰物质的假阴性或阳性结果；目标3：我们将评估灵敏度 和特异性，以及我们完全集成系统的正面和负面预测值（合并的 与商业NAAT相比，带有检测模块的样品制备模块。到阶段结束 ii，我们将被毒死以制造和使用我们的快速（<35分钟），廉价，用户友好，无仪器， 在美国进行临床试验的敏感和特定的GC POC检验，以获得FDA调节清除率。我们的整体 目的是部署我们的GC POC诊断，以用于医生办公室，小城市和乡村诊所，青少年， 家庭和性病诊所，ERS和医院诊所，其他测试网站以及资源约束的设置 世界。