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Updated: Feb 17, 2026
Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
Published on: September 25, 2018
Detection of simultaneous multi-mutations using base-quenched probe
Huihui Mao1, Guanghua Luo1, Jun Zhang1
1Comprehensive Laboratory, Changzhou Key Lab of Individualized Diagnosis and Treatment Associated with High Technology Research, The Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
Abstract:
The base-quenched probe method for detecting single nucleotide polymorphisms (SNPs) relies on real-time PCR and melting-curve approaches. Here, we applied the most common commercial fluorophores including FAM, HEX, CY5, CY3, TET, JOE, Texas Red and ROX for labeling probes to detect multi-mutations simultaneously according to the different fluorescence channels. Accuracy of the method was confirmed by direct sequencing. The results demonstrated that all above dyes could be influenced by bases and could be applied to detect SNPs. Furthermore, this method was applied to detect APOM rs707921, APOM rs707922 and MCP-1 rs1024611 simultaneously, which was demonstrated successfully.
