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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的沙眼衣原体 (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 亿病例，2014 年美国约有 144 万病例。大多数女性和男性感染无症状，促进不受控制的传播，可能导致盆腔炎、不孕症、慢性盆腔疼痛和危及生命的宫外孕。 Ct 也是侵袭性鳞状细胞癌的危险因素子宫颈癌和 HIV-1 感染的复杂因素。患者经常在没有经过治疗的情况下接受治疗明确诊断，这可能导致抗生素使用不当并可能导致耐药性。主要阻止 Ct 感染的一个障碍是缺乏护理点 (POC) 诊断来提高早期发现率减少感染率和后遗症。当前的 CT 诊断依赖于商业核酸扩增测试（NAAT）的敏感性和特异性各不相同，同一样本通常缺乏一致的结果类型。 NAAT 价格昂贵，需要设备和训练有素的操作员，需要几天的时间才能得到结果，并且可能会导致失访和延误治疗。因此，当前的 NAAT 不适合 POC 诊断。我们的团队由 Ct STD 和 POC 开发专家 Deborah Dean 博士以及初创公司 Diassess 组成拥有 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 核酸原型样品制备模块中无需仪器，5 分钟内即可从宫颈内拭子中提取酸； 2）拥有无需仪器的多重检测模块，可对 Ct 核酸进行 30 分钟比色检测； 3）拥有经过验证的检测方法，可检测所有 Ct 参考菌株，区分性病淋巴肉芽肿 (LGV) 和非 LGV 毒株并检测人类 DNA； 4) 已证明检测结果符合标准 200 份残留宫颈内膜患者样本的 NAAT 结果。在第二阶段，我们将推进第一阶段的结果。目标 1：利用不断扩大的参考和临床 Ct 基因组序列集合，完善我们的引物，根据需要更换失败的引物，并确保我们改进的 Ct 引物扩增检测能够检测到 Ct 参考和多种 Ct 临床菌株，不与性传播病原体发生交叉反应，常见的阴道/宫颈种类；目标 2：优化样品制备化学、扩增测定设计阴道、尿道和宫颈内拭子以及干扰物质的比色化学；目标 3：评估我们完全集成的系统（组合的样品制备模块）的灵敏度和特异性与商业 NAAT 相比。到第二阶段结束时，我们将做好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、灵敏且特异的 Ct 在美国进行 POC 临床试验测试，通过 510(k) 途径获得 FDA 监管许可。我们的整体我们的目标是部署我们的 Ct POC 诊断，供医生办公室、小到大城市和农村诊所、青少年和儿童使用。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。