Ctnnb1 mutation detection kit, method, and use in hcc detection and management

A kit for characterizing, in a biological sample containing at least one first allele of CTNNB1, at least one second allele in a genomic region of CTNNB1, the kit comprising:

• a first pair of primers, configured to specifically bind sequences flanking the genomic region to thereby allow amplification of at least one polynucleotide harboring the genomic region in a first PCR reaction;
• and
• at least one clamp, each configured to bind to one of the at least one first allele but not any of the at least one second allele at an annealing temperature in the first PCR reaction to thereby selectively suppress amplification of the one of the at least one first allele but still allow amplification of the at least one second allele;
• wherein:
• at least one of the first pair of primers comprises an oligonucleotide of an artificial sequence at a 5′-end thereof, configured to interrupt a secondary structure of DNA molecules of CTNNB1 or to increase a Tm of the at least one of the first pair of primers in the first PCR reaction using amplified products as templates to thereby increase an efficiency of the amplification of the at least one polynucleotide in the first PCR reaction.