Kitchen Range Oven Enabled One-tube RT-LAMP for RNA Detection at Home - A Potential Solution for Large-scale Screening of COVID-19

Abstract

Rapid, low-cost, and user-friendly molecular diagnostic methods are prerequisite to address the outbreaks of infectious diseases. Especially during the outbreak of COVID-19 pandemic, there is an urgent need to build the global testing capacity up to 100-fold above what is achievable with current standard approaches. However, current gold standard methods such as RT-PCR and isothermal PCR (e.g., RT-LAMP), which are routinely conducted in laboratories, suffer from limit capacity due to the requirement of special equipment, multiple sample handling steps, and/or the need of skilled personnel. In this study, a kitchen Range Oven enabled RT-LAMP was conducted at a residential home without sacrificing the performance in RNA detection. In addition, one-step, closed-tube RT-LAMP for the detection of target RNA in the oven was accomplished by pre-loading sample collection solution and RT-LAMP reaction reagents into the bottom and cap cavity of PCR tube, respectively. After the addition of target RNA into sample collection solution through either swab swirling or direct pipetting, a flip-and-swing enabled mixing of sample solution and RT-LAMP reaction reagents was conducted first, followed by RT-LAMP detection at a constant temperature in pre-heated oven. The RNA positive sample can be obviously differentiated from the RNA negative sample through both naked-eye based turbidity detection and fluorescence detection under UV light. In conjunction with the user-friendly one-step, closed-tube concept, this study indicates that it is feasible to run RT-LAMP at home using oven with minimum involvement of end-users, thus offering an excellent molecular detection platform, which has the potential to boost the detection capacity for any nucleic acid target. It could be a potential solution for large-scale screening of COVID-19 in short time as the tests can be conducted by residents at home, without the need of well-trained health caregivers and capacity limitation.