Methods for rapidly detecting single or multiple sequence alleles in a
sample nucleic acid are described. Provided are all of the
oligonucleotide pairs capable of annealing specifically to a target
allele and discriminating among possible sequences thereof, and ligating
to each other to form an oligonucleotide complex when a particular
sequence feature is present (or, alternatively, absent) in the sample
nucleic acid. The design of each oligonucleotide pair permits the
subsequent high-level PCR amplification of a specific amplicon when the
oligonucleotide complex is formed, but not when the oligonucleotide
complex is not formed. The presence or absence of the specific amplicon
is used to detect the allele. Detection of the specific amplicon may be
achieved using a variety of methods well known in the art, including
without limitation, oligonucleotide capture onto DNA chips or
microarrays, oligonucleotide capture onto beads or microspheres,
electrophoresis, and mass spectrometry. Various labels and
address-capture tags may be employed in the amplicon detection step of
multiplexed assays, as further described herein.