Tetrahedron supported click ligation initiated by dual recognition for precise bacterial analysis

For the 16S rRNA gene of bacterial analysis, the current usage of single recognition probe always causes the false positive result. Meanwhile, it is usually impossible for direct ligation of two free DNA strands modified with click ligation groups in the solution. In our work, A DNA tetrahedron supported click ligation has been elaborately designed; thereby a new method has been further developed for bacterial analysis with dual recognition on two target regions of 16S rRNA gene. Compared with free click ligation, DNA tetrahedron supported click ligation exhibits high reaction rate and ligation efficiency as a result of proximity effect on the supporting interface. The designed DNA tetrahedron can simultaneously bind with two target regions of 16S rRNA gene in bacteria, inducing the proximity of reaction groups and efficient occurrence of click ligation. The established method shows the practical applicability in the serum sample. In a word, inspired by high ligation efficiency on the interface, DNA tetrahedron supported click ligation has been firstly developed and served for bacterial analysis through dual recognition with high specificity, high sensitivity and good performance.