Multiplexed Detection of Mosquito-Borne Viruses at the Point-of-Care

在护理点对蚊媒病毒进行多重检测

• 批准号: 10560649
• 负责人: Z. Hugh Fan
• 金额: $ 37.19万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-11 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560649
• 关键词: Ache; Address; Affect; Anxiety; Arboviruses; Area; Arthralgia; Authorization documentation; Bacterial Infections; Benchmarking; Biological Assay; Blood; Blood specimen; Brazil; Caribbean region; Centers for Disease Control and Prevention (U.S.); Chikungunya virus; Clinic; Clinical; Clinical Management; Clinics and Hospitals; Coin; Country; Cytolysis; DNA Primers; Dengue; Dengue Infection; Dengue Shock Syndrome; Dengue Virus; Detection; Devices; Diagnosis; Diagnostic; Disease Outbreaks; Enzyme-Linked Immunosorbent Assay; Epidemic; Exanthema; Fever; Florida; General Population; Genetic; Goals; Haiti; Headache; Human; Human Resources; Infant; Infection; Laboratories; Latin America; Life; Link; Mayaro virus; Mediating; Medical; Methods; Microcephaly; Monitor; Mothers; Neurologic Deficit; Newborn Infant; Nucleic Acids; Paper; Patients; Performance; Polymerase Chain Reaction; Population; Pregnancy; Preparation; Process; Public Health; RNA; RNA Viruses; RNA amplification; Recommendation; Reporting; Research; Reverse Transcription; Saliva; Sampling; Serotyping; Specificity; Symptoms; Syndrome; Testing; Urine; Virus; Virus Diseases; World Health Organization; ZIKV infection; Zika Virus; amplification device; authority; chikungunya infection; clinical care; cost; design; detection platform; detection sensitivity; diagnostic platform; disability; epidemic potential; improved; innovation; interest; isothermal amplification; joint inflammation; mosquito-borne; multiplex detection; operation; pathogen; point of care; point of care testing; point-of-care diagnostics; population based; saliva sample; screening; vector mosquito; viral detection; viral transmission; virtual

Project Summary Dengue virus (DENV), Zika virus (ZIKV), Chikungunya virus (CHIKV), and Mayaro virus (MAYV) are all mosquito-borne RNA viruses. They are public health concerns because (1) DENV and CHIKV cause hundreds of millions of infections each year, with significant burdens in affected areas, (2) the outbreak of ZIKV in Brazil in 2015/2016 caused anxieties to general population due to its association with microcephaly of newborns, and (3) MAYV emerged in Central and Southern America recently and has the potential for epidemic spread. Because these virus infections have virtually identical clinical presentation and they often circulate concurrently, it is important to have a point-of-care (POC) testing platform to accurately identify virus infection for clinical management of patients, including different complications from these viruses. According to the Centers for Disease Control and Prevention (CDC), the current methods authorized for assessing DENV, ZIKV, and CHIKV infections include reverse transcription polymerase chain reaction (RT- PCR) assay and enzyme-linked immunosorbent assay (ELISA). However, these assays are carried out in laboratories, not at POC in a clinic or in an infected field. It is also important to note that virus infection can cause asymptomatic infections, up to 80% of ZIKV patients, 50% of DENV patients, and 28% of CHIKV patients, respectively. As a result, POC testing in the field will be more valuable for screening asymptomatic patients and monitoring possible virus transmission than a laboratory test because only symptomatic patients go to hospitals or clinics for seeking medical help or to be screened. To address the need, we propose to develop a POC diagnostic platform called Valve-enabled Lysis, paper- based RNA Enrichment, and RNA Amplification Devices (VLEAD). VLEAD will integrate sample preparation— including virus lysis and RNA enrichment—with nucleic acid amplification for simultaneous detection of these viruses. To achieve the goal, we aim to (1) develop multiplexed VLEAD for simultaneous detection of ZIKV, DENV, CHIKV and MAYV; (2) optimize VLEAD using various samples and compare the suitability of the device for urine, saliva and blood samples; and (3) validate VLEAD using clinical samples and compare VLEAD with the benchmark methods including conventional RT-PCR. The significance of the research lies in the following aspects. First, these mosquito-borne RNA viruses are a public health concern. An accurate and sample-to-answer virus detection platform at POC will be useful for clinical care and patient management. Second, a large percentage of these virus infections are asymptomatic, thus a non-invasive POC platform would be very beneficial for screening the general population in the infected area and monitoring virus transmission. Third, the VLEAD platform can be adapted for detecting other pathogens of interest, with a potential to have more societal impacts.

项目概要 登革热病毒（DENV）、寨卡病毒（ZIKV）、基孔肯雅病毒（CHIKV）和马亚罗病毒（MAYV）都是 蚊媒RNA病毒。它们是公共卫生问题，因为 (1) DENV 和 CHIKV 导致数百万人 每年有数百万感染者，给受影响地区带来巨大负担，(2) 巴西寨卡病毒爆发 2015/2016 年，由于与新生儿小头畸形相关，引起了普通民众的焦虑，以及 （3）MAYV近期在中南美洲出现，具有疫情传播的潜力。 因为这些病毒感染具有几乎相同的临床表现并且经常传播 同时，拥有一个即时检测（POC）平台来准确识别病毒感染也很重要 用于患者的临床管理，包括这些病毒引起的不同并发症。 根据美国疾病控制与预防中心 (CDC) 的说法，目前授权的方法 评估 DENV、ZIKV 和 CHIKV 感染包括逆转录聚合酶链反应 (RT- PCR）测定和酶联免疫吸附测定（ELISA）。然而，这些测定是在 实验室，而不是诊所或受感染区域的 POC。还需要注意的是，病毒感染可以 导致无症状感染，高达 80% 的 ZIKV 患者、50% 的 DENV 患者和 28% 的 CHIKV 患者 分别为患者。因此，现场 POC 检测对于筛查无症状感染者将更有价值 与实验室测试相比，监测可能的病毒传播，因为只有有症状的患者 前往医院或诊所寻求医疗帮助或接受检查。 为了满足这一需求，我们建议开发一个名为 Valve-enabled Lysis 的 POC 诊断平台，论文 - 基于 RNA 富集和 RNA 扩增设备 (VLEAD)。 VLEAD 将整合样品制备—— 包括病毒裂解和 RNA 富集——通过核酸扩增同时检测这些 病毒。为了实现这一目标，我们的目标是 (1) 开发用于同时检测 ZIKV 的多重 VLEAD， DENV、CHIKV 和 MAYV； (2) 使用各种样本优化VLEAD并比较器件的适用性 用于尿液、唾液和血液样本； (3) 使用临床样本验证 VLEAD 并将 VLEAD 与 基准方法包括常规RT-PCR。 研究的意义在于以下几个方面。首先，这些蚊媒RNA病毒是 公共卫生问题。准确且从样本到答案的 POC 病毒检测平台将有助于 临床护理和患者管理。其次，这些病毒感染者中有很大一部分是无症状的， 因此，非侵入性 POC 平台将非常有利于筛查感染者的一般人群 区域和监测病毒传播。第三，VLEAD平台可以适用于检测其他 感兴趣的病原体，有可能产生更多的社会影响。