Ultra‐Fast Single‐Nucleotide‐Variation Detection Enabled by Argonaute‐Mediated Transistor Platform

Abstract “Test‐and‐go” single‐nucleotide variation (SNV) detection within several minutes remains challenging, especially in low‐abundance samples, since existing methods face a trade‐off between sensitivity and testing speed. Sensitive detection usually relies on complex and time‐consuming nucleic acid amplification or sequencing. Here, a graphene field‐effect transistor (GFET) platform mediated by Argonaute protein that enables rapid, sensitive, and specific SNV detection is developed. The Argonaute protein provides a nanoscale binding channel to preorganize the DNA probe, accelerating target binding and rapidly recognizing SNVs with single‐nucleotide resolution in unamplified tumor‐associated microRNA, circulating tumor DNA, virus RNA, and reverse transcribed cDNA when a mismatch occurs in the seed region. An integrated microchip simultaneously detects multiple SNVs in agreement with sequencing results within 5 min, achieving the fastest SNV detection in a “test‐and‐go” manner without the requirement of nucleic acid extraction, reverse transcription, and amplification.