A Rapid, Low-Cost Point of Care Diagnostic for detection of Zika virus RNA

用于检测寨卡病毒 RNA 的快速、低成本护理点诊断

• 批准号: 9255921
• 负责人: Sahar Usmani-Brown
• 金额: $ 22.5万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9255921
• 关键词: Acute; Adult; Aedes; African; Amniocentesis; Area; Asians; Bathing; Biological Assay; Blood; Blood specimen; Caribbean region; Central America; Chikungunya virus; Clinical; Colombia; Communities; Complex; Congenital Abnormality; Culicidae; Dengue; Dengue Virus; Dependence; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Disease Outbreaks; Distant; Epidemic; Epidemiologic Monitoring; Equipment; Evaluation; Exanthema; Eye; Female of child bearing age; Fetus; Fever; Flavivirus; Goals; Guillain-Barré Syndrome; Health; Hour; Human; Imagery; Laboratories; Lateral; Mediating; Methods; Microcephaly; Micronesia; Mothers; Newborn Infant; Nucleic Acids; Patients; Performance; Phase; Pregnancy; Pregnant Women; Prenatal care; Procedures; Public Health; Puerto Rico; Quantitative Reverse Transcriptase PCR; RNA; RNA Sequences; RNA-Directed DNA Polymerase; Reaction; Reporting; Research; Resources; Risk; SYBR Green I; Saliva; Sampling; Sensitivity and Specificity; Serologic tests; Serum; Small Business Innovation Research Grant; South America; Southeastern United States; Specificity; Specimen; System; Techniques; Technology; Temperature; Testing; Time; TimeLine; Ultrasonography; United States Dept. of Health and Human Services; Urine; Viral; Viral Genome; Virus; Virus Diseases; Water; West Nile virus; Woman; Work; Zika Virus; assay development; authority; base; chikungunya; clinical Diagnosis; clinical diagnostics; cost; cost effective; cross reactivity; design; diagnostic assay; experience; point of care; point-of-care diagnostics; pregnant; prenatal; prototype; sample collection; screening; transmission process; validation studies; viral RNA; viral detection

ABSTRACT Zika virus (ZIKV) has rapidly emerged and spread through South and Central America, the Caribbean, and Puerto Rico since its last outbreak in Micronesia in 2007. Transmitted by the mosquito Aedes sp, its forecasted spread will have a major impact on the Southeast U.S. Most ZIKV infections remain asymptomatic or present with non-specific rash and fever; therefore, they have been difficult to diagnose and report. However, two major health consequences appear to be associated with the ZIKV outbreak which sets it apart from other flaviviruses such as West Nile Virus and Dengue; namely; (a) transmission from an infected mother to fetus resulting in reports of microcephaly in fetuses; and, (b) Guillain-Barre syndrome (GBS) in adults. Several teams have now developed qRT-PCR assays to detect ZIKV. However such tests are relatively expensive, require well-equipped laboratories with specialized equipment, and the procedure takes at least 3 hours to finish. There is an urgent need for a rapid, sensitive, specific and economical diagnostic test for ZIKV. Such an assay could be routinely used in resource-poor settings as well as in doctors' offices, including as part of regular prenatal care. Therefore, the goal of the SBIR Phase I project is to use loop mediated isothermal nucleic acid amplification (LAMP) to develop a rapid, sensitive point of care diagnostic for ZIKV. This technology is of low complexity, requiring only a water bath. Colorimetric results are visible to the naked eye in one hour or less. We will use serial dilutions of ZIKV and other viruses (including West Nile virus, Dengue viruses, and Chikungunya virus) spiked in human blood, saliva, and urine to determine the assay's sensitivity and specificity. Based on our laboratory's extensive previous experience with LAMP assay development, we forecast a lower limit of detection of 10-100 viral genomes, with very high specificity. In addition to rapid, colorimetric ZIKV detection, we will also develop a rapid, lateral flow assay to facilitate the differential analysis with other flaviviruses. The development of rapid point-of-care assays will reduce the dependence on central laboratory testing facilities for epidemiologic surveillance and clinical diagnosis, a key advantage in the resource-poor areas where the epidemic is currently prevalent. Further, we anticipate that the availability of a rapid test will result in the incorporation of ZIKV testing into the sustainable, routine evaluation of women who are pregnant or anticipating pregnancy, as well as their partners.

抽象的 寨卡病毒 (ZIKV) 已迅速出现并在南美洲、中美洲、加勒比地区和加勒比海地区传播。 波多黎各自 2007 年在密克罗尼西亚爆发上次疫情以来。预计由伊蚊传播 传播将对美国东南部产生重大影响 大多数 ZIKV 感染仍然无症状或存在 伴有非特异性皮疹和发烧；因此，它们很难诊断和报告。然而，两个 重大健康后果似乎与 ZIKV 爆发有关，这使其与其他疾病不同 黄病毒，例如西尼罗河病毒和登革热；即； (a) 从受感染的母亲传播给胎儿 导致胎儿小头畸形的报告； (b) 成人格林-巴利综合征 (GBS)。一些 团队现已开发出 qRT-PCR 检测方法来检测 ZIKV。然而这样的测试相对昂贵， 需要设备齐全的实验室和专门的设备，整个过程至少需要3个小时 结束。迫切需要一种快速、灵敏、特异且经济的 ZIKV 诊断测试。 这种测定可以在资源匮乏的环境以及医生办公室中常规使用，包括作为 定期产前护理。因此，SBIR一期项目的目标是利用环介导等温 核酸扩增 (LAMP) 开发快速、灵敏的 ZIKV 护理诊断方法。这 技术复杂度低，仅需要水浴。比色结果肉眼可见 一小时或更短。我们将使用 ZIKV 和其他病毒（包括西尼罗河病毒、登革热病毒）的系列稀释液 病毒和基孔肯雅病毒）掺入人体血液、唾液和尿液中以确定检测的灵敏度 和特异性。基于我们实验室之前在 LAMP 检测开发方面的丰富经验，我们 预测 10-100 个病毒基因组的检测下限，具有非常高的特异性。除了快速之外， 比色 ZIKV 检测，我们还将开发一种快速侧流检测，以方便差异分析 与其他黄病毒。快速现场检测的发展将减少对中央检测的依赖 用于流行病学监测和临床诊断的实验室检测设施是该领域的一个关键优势 目前疫情流行的资源匮乏地区。此外，我们预计将有一个 快速检测将使 ZIKV 检测纳入对患有以下疾病的女性的可持续的常规评估： 已怀孕或即将怀孕的人以及他们的伴侣。