Tepnel Medical Limited

John Douglas Oultram

Conor Mulrooney

Jacqueline Clare Coutts

The present invention concerns detection of target molecules using methods of enzymatically catalysed amplification of target associated detectable structures.

Enzymatically catalysed signal amplification

The present invention relates to a method of amplifying a nucleic acid sequence present in a first strand of a double stranded nucleic acid molecule wherein said molecule incorporates an unmodified recognition site for a restriction enzyme capable of cutting the first strand at the 5&prime; end of the sequence therein to be amplified and leaving the 3&prime;-cut region of the second strand projecting beyond the cut site in the first strand. The method further comprises treating said molecule with said enzyme in the presence of a strand displacing polymerase and unmodified nucleotides for incorporation in an extending nucleic acid strand such that there is or becomes hybridised to said 3&prime;-end region of the second strand a primer sequence complementary thereto whereby said primer sequence is extended in the 5&prime; to 3&prime; direction using the second strand as a template to regenerate the restriction endonuclease cut site and displace the sequence to be amplified.

Amplification of nucleic acids

Methods for synthesizing oligonucleotide products and detecting single-stranded target nucleic acid sequences, are disclosed. The methods include amplifying a template sequence to produce a complementary strand hybridized to the template sequence, which is subsequently cleaved by at least one cleavage agent displacing the complementary strand as a single stranded sequence complementary to the template sequence.

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Stretford, United Kingdom

John Douglas Oultram