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: 10349474
• 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/10349474
• 关键词: Ache; Address; Affect; Americas; Anxiety; Arboviruses; Area; Arthralgia; Bacterial Infections; Benchmarking; Biological Assay; Blood; Blood Circulation; Blood specimen; Brazil; Caribbean region; Centers for Disease Control and Prevention (U.S.); Chikungunya virus; Clinic; Clinical; Clinical Management; 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; Hospitals; 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; Patient Monitoring; Patients; Performance; Polymerase Chain Reaction; Population; Pregnancy; Preparation; Process; Public Health; RNA; RNA Viruses; RNA amplification; Reporting; Research; Reverse Transcription; Saliva; Sampling; Sensitivity and Specificity; Serotyping; Symptoms; Syndrome; Testing; Time; Urine; Virus; Virus Diseases; World Health Organization; ZIKV infection; Zika Virus; amplification device; base; chikungunya infection; clinical care; cost; design; detection platform; detection sensitivity; diagnostic platform; disability; improved; innovation; interest; isothermal amplification; joint inflammation; mosquito-borne; multiplex detection; operation; pathogen; patient screening; 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.

项目总结