Toehold-controlled ligation and transcription for accurate COVID-19 genotyping

The global pandemic of coronavirus disease 2019 (COVID-19) has significant impact on the entire human society. However, in the face of continually emerging more contagious SARS-CoV-2 variant, the risk to bog down into more severe crisis is around us anytime. Here, we introduce an isothermal, ultrasensitive method for identifying important SNV mutations of SARS-CoV-2. It is based on combined specificity of toehold-assisted linear probe ligation and in vitro transcription signal enlargement, TLT. A ready-to-use panel of TLT assay is developed including detection of 80 crucial SARS-CoV-2 SNVs, by which people could response to the next coming contagious virus variant more rapidly. These advanced point-of-care features make TLT one good approach for large scale population testing of special SARS-CoV-2 variants of interesting.