Rapid detection of disease-causing pathogens is very crucial for efficient management of communicable diseases. During a pandemic such as COVID-19, it is imperative that rapid, sensitive and simple screening technologies that can be fully integrated into near real-time medical reporting infrastructure are available, so as to speedily identify viral carriers and slow current and future viral outbreaks.
While RT-PCR is the most commonly used method for detecting pathogen-related RNA, it requires expensive thermocycling equipment and is comparatively slow. Isothermal methods promise procedural simplicity, but have traditionally suffered from amplification artifacts that tend to preclude easy identification of target nucleic acids.
We have engineered an isothermal RNA detection system called MDetect that reports the presence of RNA template by fluorogenic aptamer (RNA Mango) synthesis in a simple fluorogenic assay. As this isothermal nucleic acid detection scheme directly produces a fluorescent reporter, real-time detection is intrinsic to the assay. Nested Mango NASBA is highly specific and, in contrast to existing RNA detection systems, offers a cheap, simple and specific way to rapidly detect single molecules of pathogenic RNA.