Patent ID: 7972818

Claim:
A method for multiplexed detection of PCR amplified products comprising: providing a DNA sample having unique target DNA sequences of interest for a sample multiplex PCR amplification reaction, providing necessary reagents and primers for said sample multiplex PCR amplification reaction, wherein said reagents and primers comprise at least one unique reporting DNA sequence, each of the at least one unique reporting DNA sequence complementary to a region on a corresponding unique target DNA sequence of interest, wherein each unique reporting DNA sequence includes a fluorescence resonant energy transfer marker, forming a sample PCR product by conducting said sample multiplex PCR amplification reaction using said DNA sample, and said reagents and primers, wherein the at least one unique reporting DNA sequence is modified upon hybridization with the corresponding unique DNA target sequence, each modified unique reporting DNA sequence detectably distinguishable from a corresponding unmodified unique reporting DNA sequence, forming a microsphere mix comprising a plurality of encoded microspheres, each microsphere having a unique physically-detectable code selected from a number of unique physically-detectable codes detectable by flow cytometry, each microsphere bound to a unique biotinylated oligonucleotide, wherein each unique physically-detectable code corresponds to specific oligonucleotide having a DNA sequence able to hybridize a corresponding unmodified unique reporting DNA sequence forming a sample hybridization product by adding said sample PCR product to said microsphere mix, forming a background PCR product by conducting a background multiplex PCR amplification reaction using at least one control DNA sequence, the control DNA sequence differing from each of the unique target DNA sequences of interest, wherein said background PCR amplification reaction is run under the same conditions and using the same primers and reagents as said sample PCR amplification reaction, forming a background hybridization product by adding said background PCR product to said microsphere mix, wherein the presence or absence of false positives is determined; and determining the existence of a number of detected target DNA sequences of interest specified by said reporting DNA sequence by comparing the fluorescence of said sample hybridization product with said background hybridization product using flow cytometry, wherein said number of detected target DNA sequences of interest is not higher than said number of unique physically-detectable codes detectable by flow cytometry.