Respriatory Virus Infections: Rapid Low Cost Molecular Detection

描述（由申请人提供）：在发展中国家，急性呼吸道感染（ARI）每年导致约190万儿童死亡，其中20%在印度。在印度，因急性呼吸道感染住院的儿童中有45%发现了病毒。许多分子诊断工具的费用限制了它们在发展中国家的使用。逆转录环介导等温扩增（LAMP）技术是一种特异性高、效率高、成本低、快速扩增RNA的新技术。目的1。开发一种敏感、特异、低成本的呼吸道病毒分子诊断方法。将建立呼吸道合胞病毒（RSV）、偏肺病毒（MPV）和副流感病毒（PIV）的LAMP检测方法。将评估LAMP检测的视觉和机器检测。将开发LAMP定量实时检测方法。目标2。评估LAMP在临床样本中检测呼吸道病毒的敏感性、特异性和成本效益。灯法将与RT-PCR法进行病毒检测比较。将使用的样本来自德里现有的一组独特的临床样本，包括从儿童急性呼吸道感染研究中前瞻性收集的样本。定量实时LAMP将用于检验病毒载量与疾病严重程度相关的假设。该提案响应PAR-07-216，印度-美国母婴健康和人类发展研究方案。这是一个涉及美国和印度研究人员的合作项目，旨在加强儿童健康、疾病预防、产品开发和技术转让。目标是将新的分子检测技术转化为快速呼吸道病毒诊断的实用工具。拟议的研究与公共卫生有关，因为呼吸道病毒感染是发达国家和发展中国家儿童疾病的一个重要原因。改进的病毒检测方法可以提高呼吸道病毒的诊断，支持呼吸道病毒发病机制的研究以及抗病毒药物和疫苗的开发。

DESCRIPTION (provided by applicant): Acute respiratory infections (ARI) result in ~ 1.9 million childhood deaths per year in developing countries, 20% of the deaths are in India. Viruses were found in 45 % of children hospitalized with ARI in India. The expense of many molecular diagnostic tools has limited their use in developing countries. Reverse-transcription loop mediated isothermal amplification (LAMP) is a novel technique that can amplify RNA with high specificity, efficiency, and rapidity at a low cost. Aim 1. Develop a sensitive, specific, low-cost molecular diagnostic assay for respiratory virus detection. LAMP assays will be established for respiratory syncytial virus (RSV), metapneumovirus (MPV), and parainfluenza virus (PIV). Visual and machine detection for LAMP assays will be evaluated. Quantitative real-time LAMP assays will be developed. Aim 2. Evaluate the sensitivity, specificity and cost-effectiveness of LAMP for the detection of respiratory viruses in clinical samples. Lamp assays will be compared to RT-PCR for virus detection. The samples to be used are from a unique collection of clinical samples available in Delhi, including prospectively collected samples from studies of ARI in children. Quantitative real-time LAMP will be used to test the hypothesis that viral loads correlate with disease severity. This proposal is responsive to PAR-07-216, Indo-US Program on Maternal and Child Health and Human Development Research. It is a collaborative project involving U.S. and Indian investigators to enhance child health, disease prevention, product development and technology transfer. The goal is to translate new molecular detection techniques into practical tools for rapid respiratory virus diagnosis. The proposed studies are relevant to public health because respiratory virus infections are an important cause of disease among children in both developed and developing countries. Improved virus detection methods can improve respiratory virus diagnosis and support studies of respiratory virus pathogenesis along with antiviral and vaccine development.