AMPLIFICATION OF TARGET NUCLEIC ACIDS USING GAP FILLING LIGASE CHAIN REACTION

摘要

2111962 9300447 PCTABS00019 An improved "gap filling" embodiment of the Ligase Chain Reaction (LCR) is described. Gap filling LCR is LCR wherein at least one of the probes is recessed so that a gap is formed between the adjacent probes when they are hybridized to target. The gap is filled using polymerase and deoxyribonucleotide triphosphates before ligation of the probes together. There are single and double gap versions, depending on whether one or two probes are recessed and require filling before ligation. The improvement resides in selecting and using target sequences such that only a single type, or two types, of deoxyribonucleotide triphosphate(s) are required to fill double gaps each being 1-10 bases in length, preferably 1-3 bases. Probes having specific sequences are claimed for a number of pathogens.