Patent ID: 6537783
Filing Date: 2003-03-25
Classification: C07H,C12N,C12P,C12Q

Abstract:
Process for amplifying a sequence of a target nucleic acid which is present in a sample, according to which:at least the following are available: the sequence of a target nucleic acid, at least one oligonucleotide primer which is specific for the target sequence, one or more enzyme activities that amplify the sequence and nucleotides, and the target sequence is amplified under conditions which are suitable for the enzymic activity or activities, characterized in that, among the nucleotides available, at least one nucleotide is a prefunctionalized nucleotide which differs from the other nucleotides at least by the presence of at least one unprotected covalently reactive function which is arranged in at least one predetermined site of the base of said nucleotide and which is an electrophilic organic chemical function selected from the group consisting of aldehyde, activated ester, carboxylic acid, isothiocyanate, haloacyl derivatives and sulfonyl chloride functions or a nucleophilic organic chemical function selected from the group consisting of thiol, oxyamino, alkoxyamino, hydrazine and hydrazide functions, and in that, in addition the resulting prefunctionalized amplification product is reacted, directly or indirectly, with a reagent which contains a covalently anti-reactive function which is specific for the reactive function of the prefunctionalized nucleotide and a functional labeling group, with this latter being a non-protein, in order to obtain a functionalized amplification product, wherein one of (1) the reactive function of said prefunctionalized nucleotide and (2) the anti-reactive function of said reagent is an aldehyde function.