Development of a rapid multiplex CRISPR-based testing pathway for tuberculosis and COVID-19

项目摘要 结核病和新冠肺炎是目前世界上最致命的病原体， 每年有140万人死于结核病和250万人死于新冠肺炎，两者都是由于潜在的传播而持续存在的 未确诊的、无症状的感染。在许多结核病-冠状病毒共同流行的人群中 在亚洲、撒哈拉以南非洲、南美洲和东欧，迫切需要 对结核病和冠状病毒进行广泛、主动、症状不可知的筛查，以控制传播和 降低发病率和死亡率。然而，COVID和TB的同时负担构成了 在带宽和资源严重有限的情况下，这些医疗保健系统面临巨大压力 积极发现和监测病例。因此，大约有40万人死于结核病 预计未来5年与前几年相比，这是新冠肺炎的直接后果。 除了它们在全球的共同流行率和无症状传播的可能性之外，结核病和CoVID 临床症状和体征重叠、危险因素和共用呼吸道传播 途径允许一种组合的快速筛查方法，该方法可以检测到这两种感染 一个样本和测试路径。这将使i)更广泛的筛查，ii)在更高 效率--只需对更少的人进行筛查即可检测到一名结核病或柯萨奇病毒感染者。 我们建议利用我们现有的结核病和冠状病毒非侵入性样本(例如，痰， 浓缩唾液、口腔拭子)、常用处理方法和一种新的护理点 兼容CRISPR-CAS 13 COVID诊断系统(SIRE)，用于试验简化方法 同时筛查结核病和柯萨奇病毒病。具体地说，为了开发这种联合筛查 战略，我们将追求以下目标：1)过渡到一个护理点CRISPR平台，用于 结核病和冠状病毒的多重筛查，以及2)优化结核和冠状病毒的共提取方法 痰和痰标本。这些相辅相成的目标将为 实现对COVID和TB的简化测试和控制，并适用于快速、 对未来大流行进行多途径筛查和监测。

PROJECT ABSTRACT Tuberculosis (TB) and COVID-19 (COVID) are currently the deadliest pathogens worldwide, with 1.4 million TB and 2.5 million COVID deaths annually, both perpetuated by potential transmission from undiagnosed, asymptomatic infection. In the many TB-COVID co-endemic populations in Asia, sub-Saharan Africa, South America, and Eastern Europe, there is a critical need for widespread, active, symptom-agnostic screening of TB and COVID to control transmission and reduce morbidity and mortality. However, the simultaneous burden of COVID and TB poses enormous stress on these health care systems with severely limited bandwidth and resources for active case finding and surveillance. Consequently, around 400 thousand more TB deaths are estimated in the next 5 years compared to prior years as a direct consequence of COVID. Beyond their global co-prevalence and potential for asymptomatic transmission, TB and COVID’s overlapping clinical signs and symptoms, risk factors, and shared respiratory transmission pathways allow for a combined rapid screening approach that can detect both infections using one sample and testing pathway. This would enable i) more wide-spread screening, ii) at higher efficiency – fewer individuals need to be screened to detect one TB or COVID infected individual. We propose to leverage our existing TB and COVID non-invasive samples (eg. sputum, concentrated saliva, oral swabs), common processing methods and a novel point-of-care compatible CRISPR-Cas 13 COVID diagnostic system (SHINE) to pilot a streamlined approach to simultaneously screen for TB and COVID. Specifically, to develop this combined screening strategy, we will pursue the following aims: 1) transition a point-of-care CRISPR platform for multiplex screening of TB and COVID, and 2) optimize co-extraction methods from TB and COVID sputum and sputum specimens. These complementary aims will contribute independent value to enable streamlined testing and control of both COVID and TB, and are adaptable towards rapid, multiplex screening and surveillance of future pandemics.