Determination of low-abundance single-base point mutations based on endonuclease IV and branch migration system

Combining endonuclease IV and branch migration competition systems, we have designed a new single-base mutation detection method which is capable of thermostatic, sensitive, simple, and cost-effective at the same time, while other probe method based on endonuclease IV hardly achieve. Our method has better discrimination factors (6.81–83.10) and a low abundance detection limit of mutant-type DNA (MT) (0.05% mutant-type DNA/total DNA) without complex temperature optimize process. The concentration detection limit of MT is 0.03 nM.The abundance detection limit for EGFR L858R (0.1%) and PTEN R130Q (0.05%) in clinical samples suggested that the method has potential applications in clinical diagnosis. [Display omitted] •The method combined the Endo IV and branch migration system to realize point mutations detection under constant temperature.•This method is simple, economical, and fast. It can be used to detect synthetic DNA strand and clinical samples with high sensitivity.•The method can be used to detect EGFRL858R and PTEN R130Q, they are common gene mutations cancer.