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

项目摘要 结核病（TB）和COVID-19（COVID）是目前全球最致命的病原体， 1.4每年有250万人死于结核病，250万人死于新冠肺炎， 来自未确诊的无症状感染者。在许多结核病和新冠肺炎并存的人群中， 亚洲、撒哈拉以南非洲、南美洲和东欧， 对结核病和新冠肺炎进行广泛、积极、不确定的筛查，以控制传播， 降低发病率和死亡率。然而，COVID和结核病同时造成的负担 这些医疗保健系统面临巨大压力，带宽和资源严重有限， 积极开展病例发现和监测。因此，大约40万多结核病死亡人数 估计未来五年与过往年度相比，因COVID的直接后果。 除了结核病和新冠肺炎的全球共同流行和无症状传播的可能性之外， 重叠的临床体征和症状、风险因素和共同的呼吸道传播 途径允许联合快速筛查方法，可以检测两种感染， 一个样本和测试路径。这将使i）更广泛的筛选，ii）在更高的 效率-更少的人需要进行筛查，以检测一个结核病或COVID感染者。 我们建议利用我们现有的结核病和新冠肺炎非侵入性样本（例如：痰， 浓缩唾液、口腔拭子）、常见的处理方法和新的即时护理 兼容CRISPR-Cas 13 COVID诊断系统（SHINE），以试行简化方法 同时筛查结核病和新冠肺炎具体地说，要发展这种联合筛查， 我们将追求以下目标：1）将床旁CRISPR平台过渡到 TB和COVID的多重筛选，以及2）优化TB和COVID的共提取方法 痰液和痰液标本。这些互补的目标将为以下方面提供独立的价值： 能够简化COVID和TB的测试和控制，并可适应快速， 多重筛查和监测未来的大流行病。

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.