| 摘要 |
A method for the detection of a target nucleic acid sequence in a sample, which comprises: (i) contacting the sample, which has been treated to bring any nucleic acids present into single-stranded form, (a) with a linear single stranded vector-probe capable of transforming competent bacteria when in circular form, said vector-probe comprising a nucleic acid sequence complementary to the target nucleic acid sequence, said nucleic acid sequence being present in two parts, one part of said nucleic acid being present at the 5'-end of the vector-probe, the other part being present at the 3'-end, said vector-probe also comprising a region encoding a selectable or detectable marker, under conditions that permit hybridisation of any target sequence present in the sample to the complementary sequence, and (b) with an enzyme capable of repairing a single-stranded break in a double-stranded nucleic acid structure; (ii) inactivating the repair enzyme; (iii) separating any resulting circularized single stranded linear vector-probe from the annealed target; (iv) contacting the circularized single stranded linear vector-probe, optionally in the form of the reaction mixture resulting from step (ii), with competent host bacteria that lack the marker present in the vector-probe under transformation conditions; (v) culturing the resulting transformed bacteria and investigating the presence of the marker; and a vector for use in the method. The method may also be used for the detection and/or identification of mutations in a target nucleic acid sequence. |
