Multiplex methylation-specific amplification systems and methods

Abstract

The present invention relates to systems and methods for performing multiplex amplification reactions. In particular, the present invention relates to multiplex methylation-specific amplification systems and methods.

We claim: 
     
         1 . A kit, comprising:
 a) a plurality of primer pairs that amplify a specific target nucleic acid wherein said specific target nucleic acid is a single gene or an untranslated region of a single gene, wherein said plurality of primer pairs amplifies a plurality of different nucleic acid segments within said specific target nucleic acid to generate a plurality of different amplicons;   b) a plurality of different nucleic acid probes that specifically bind to amplicons amplified by said plurality of primer pairs that amplify said specific target nucleic acid, wherein said different nucleic acid probes comprise a detectable label that is the same label in each of said plurality of different nucleic acid probes; and   c) a methylation specific detection reagent.   
     
     
         2 . The kit of  claim 1 , wherein said methylation specific detection reagent is bisulfite. 
     
     
         3 . The kit of  claim 1 , wherein said target nucleic acid is a promoter region of a gene. 
     
     
         4 . The kit of  claim 1 , wherein said target nucleic acid is GC-rich. 
     
     
         5 . The kit of  claim 1 , wherein said target nucleic acid comprises CpG islands. 
     
     
         6 . The kit of  claim 1 , further comprising one or more of a polymerase, a deoxynucleotide, a buffer, a blocking oligonucleotide, dimethyl sulfoxide (DMSO), Bovine Serum Albumin (BSA), a detergent, a solid support, a control, and a salt. 
     
     
         7 . The kit of  claim 1 , wherein said detectable label is a fluorescent label. 
     
     
         8 . The kit of  claim 1 , wherein said at least one of said plurality of different nucleic acid probes is dual labeled with a reporter and a quencher molecule.