| 摘要 |
Particular aspects provide nucleic acid amplification and detection methods comprising: providing a reaction mixture containing a target nucleic acid with an amplifiable target sequence, forward and reverse external nick-directing primers (ND-primers), at least one internal ND-primer, a strand-displacing DNA polymerase, a nick-directing endonuclease for strand-specific cleavage of ND-primer-extension products, and deoxynucleoside 5′-triphosphates; and incubating the reaction mixture with reagents, and under conditions suitable to provide for amplification of the amplifiable target sequence, wherein the amplification comprises primer extension, by least one internal ND-primer, of an external ND-primer extension product comprising the amplifiable target sequence or a portion thereof but lacking the respective external ND-primer sequence or a portion thereof. Preferably, amplification comprises using a plurality of internal ND-primers, extension of one internal ND-primer extension product by a different internal ND-primer, and amplification is isothermal and synergistic with respect to the number of primers employed. Amplification and detection kits are provided. |
