Low-Cost Instrument-free Point-of-care Platform for Multiplexed Chlamydia Diagnostics

用于多重衣原体诊断的低成本无仪器即时护理平台

• 批准号: 9202973
• 负责人: DEBORAH Anne DEAN
• 金额: $ 70.79万
• 依托单位: LUCIRA HEALTH, INC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9202973
• 关键词: Accident and Emergency department; Antibiotics; Bacterial Sexually Transmitted Diseases; Bedside Testings; Biological Assay; Birth; Blood; Cervical; Cervical Squamous Cell Carcinoma; Chemistry; Chlamydia; Chlamydia trachomatis; Cities; Clinic; Clinical; Clinical Trials; Computer Simulation; Conjunctivitis; Detection; Development; Diagnosis; Diagnostic; Drug resistance; Early Diagnosis; Ectopic Pregnancy; Ensure; Epidemic; Equipment; Female; Genomics; Goals; HIV-1; Head; Infant; Infection; Infertility; Lead; Leukocytes; Life; Lymphogranuloma Venereum; Medical; Modification; Mucous body substance; Multi-Institutional Clinical Trial; Nucleic Acid Amplification Tests; Nucleic Acids; Pathway interactions; Patients; Pelvic Inflammatory Disease; Phase; Pneumonia; Predictive Value; Preparation; Public Health; Resources; Risk Factors; Rural; Sampling; Seminal fluid; Sensitivity and Specificity; Sexually Transmitted Diseases; Site; Specificity; Specimen; Squamous cell carcinoma; Swab; System; Technology; Teenagers; Test Result; Testing; TimeLine; Training; Urethra; Vagina; Woman; abstracting; chronic pelvic pain; cost; cross reactivity; design; experience; follow-up; genome sequencing; geographic population; human DNA; instrument; male; meetings; pathogen; point of care; point-of-care diagnostics; prototype; screening; stem; transmission process; user-friendly

Abstract Chlamydia trachomatis (Ct) is the most common cause of bacterial sexually transmitted diseases (STD) with ~108 million annual cases worldwide and ~1.44 million U.S. cases in 2014. Most female and male infections are asymptomatic, facilitating unchecked transmission that can result in pelvic inflammatory disease, infertility, chronic pelvic pain, and life-threatening ectopic pregnancy. Ct is also a risk factor for invasive squamous-cell carcinoma of the cervix and a complicating factor in HIV-1 infection. Patients are often treated without a definitive diagnosis, which can lead to inappropriate antibiotic use and possibly drug resistance. The main obstacle to stemming Ct infections is the lack of a point-of-care (POC) diagnostic to increase early detection to reduce infection rates and sequelae. Current Ct diagnostics rely on commercial nucleic acid amplification tests (NAATs) that vary in sensitivity and specificity with a general lack of concordant results for the same sample type. NAATs are expensive, require equipment and highly trained operators, take a day to days for results, and can result in loss to follow up and delay in treatment. Thus, current NAATs are not suitable POC diagnostics. Our team of Dr. Deborah Dean, an expert on Ct STDs and POC development, and Diassess, a startup company with proprietary technology for POC diagnostics, showed in Phase I that we: 1) can extract Ct nucleic acids from endocervical swabs in 5 min with no instruments in a prototype Sample Preparation Module; 2) have an instrument-free multiplexed Detection Module for 30 min colorimetric detection of Ct nucleic acids; 3) have validated assays to detect all Ct reference strains, differentiate lymphogranuloma venereum (LGV) from non-LGV strains and detect human DNA; and 4) have demonstrated assay results consistent with standard NAAT results on 200 remnant endocervical patient samples. In Phase II, we will advance on Phase I results. Aim 1: Using the expanding aggregate of reference and clinical Ct genome sequences, refine our primers, replacing failed primers as needed and ensure that our refined Ct primer amplification assays detect Ct reference and diverse Ct clinical strains without cross-reactivity with sexually transmitted pathogens and common vaginal/cervical species; Aim 2: Optimize sample preparation chemistry, amplification assay design and colorimetric chemistry for vaginal, urethral and endocervical swabs, and interfering substances; Aim 3: Evaluate sensitivity and specificity 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 Ct POC test for clinical trials in the U.S. to obtain FDA regulatory clearance via the 510(k) pathway. Our overall goal is to deploy our Ct POC diagnostic for use in doctor's offices, small to large city and rural clinics, teen and STD clinics, emergency rooms, other testing sites and resource-constrained settings around the world.

摘要 沙眼衣原体（Ct）是细菌性性传播疾病（STD）的最常见原因， 全球每年约有1.08亿例，2014年美国约有144万例。大多数女性和男性感染 无症状，促进未经检查的传播，可导致盆腔炎，不孕症， 慢性盆腔疼痛和危及生命的宫外孕Ct也是浸润性鳞状细胞癌的危险因素 宫颈癌和HIV-1感染的并发因素。患者通常在没有治疗的情况下接受治疗。 明确诊断，这可能导致不适当的抗生素使用和可能的耐药性。主要 阻止Ct感染的障碍是缺乏护理点（POC）诊断，以增加早期检测， 减少感染率和后遗症。目前的Ct诊断依赖于商业核酸扩增测试 （NAAT）的灵敏度和特异性不同，同一样本通常缺乏一致的结果 类型. NAAT是昂贵的，需要设备和训练有素的操作员，需要一天到几天的结果， 可能导致失访和治疗延迟。因此，当前的NAAT不是合适的POC诊断。 我们的团队Deborah Dean博士是Ct STD和POC发展方面的专家，Diassess是一家初创公司， 一家拥有POC诊断专有技术的公司，在第一阶段表明，我们：1）可以提取Ct核酸 在原型样品制备模块中，在5分钟内从宫颈内拭子中提取酸，无仪器; 2） 具有用于Ct核酸的30分钟比色检测的无仪器多重检测模块; 3） 已经验证了检测所有Ct参考菌株的检测方法，区分性病淋巴肉芽肿（LGV）和 非LGV毒株并检测人DNA;和4）已证明检测结果与标准品一致 200例残留宫颈内患者样本的NAAT结果。在第二阶段，我们将推进第一阶段的成果。 目的1：使用参考和临床Ct基因组序列的扩展集合，改进我们的引物， 根据需要更换失败的引物，并确保我们的改良Ct引物扩增检测试剂盒检测到Ct 与性传播病原体无交叉反应性的参考和不同Ct临床菌株， 常见阴道/宫颈种属;目的2：优化样品制备化学、扩增试验设计 阴道、尿道和宫颈拭子的比色化学和干扰物质;目标3： 评估我们完全集成的系统（组合样品制备模块）的灵敏度和特异性 与商业NAAT相比，到第二阶段结束时，我们将准备 制造和使用我们的快速（<35分钟），廉价，用户友好，免仪器，灵敏和特异性Ct 在美国进行临床试验的POC测试，以通过510（k）途径获得FDA监管许可。我们的整体 我们的目标是将我们的Ct POC诊断部署在医生办公室、小型到大型城市和农村诊所、青少年和 世界各地的性病诊所、急诊室、其他检测场所和资源有限的环境。