Nucleic acid amplification primers for pcr-based clonality studies

Abstract

The invention relates to PCR-based clonality studies for among others early diagnosis of lymphoproliferative disorders. Provided is a set of nucleic acid amplification primers comprising a forward primer, or a variant thereof, and a reverse primer, or a variant thereof, capable of amplifying a rearrangement selected from the group consisting of a VH-JH IGH rearrangement, a DH-JH IGH rearrangement, a VK-JK IGK rearrangement, a VK/intron-Kde IGK rearrangement, a Vλ-Jλ IGL rearrangement, a Vβ-Jβ TCRB rearrangement, a Dβ-Jβ TCRB rearrangement, a Vγ-Jγ TCRG rearrangement, a Vδ-Jδ TCRD rearrangement, a Dδ-Dδ TCRD rearrangement, a Dδ-Jδ TCRD rearrangement, a Vδ-Dδ TCRD rearrangement, or a translocation selected from t(11;14)(BCL1-IGH) and t(14;18)(BCL2-IGH). The primers can be used in PCR-based clonality studies for early diagnosis of lymphoproliferative disorders and detection of minimal residual disease (MRD). Also provided is a kit comprising at least one set of primers of the invention.

1 . A set of nucleic amplification primers capable of amplifying a V H -J H  IGH rearrangement comprising a forward primer and a reverse primer, wherein said forward primer is selected from the V H  family primers shown in  FIG. 3B , or a variant thereof, and wherein said reverse primer is the J H  consensus primer shown in  FIG. 3B , or a variant thereof. 
     
     
         2 - 7 . (canceled) 
     
     
         8 . A set of nucleic amplification primers capable of amplifying a Vγ-Jγ TCRG rearrangement comprising a forward primer and a reverse primer, wherein said forward primer is selected from the Vγ family primers shown in  FIG. 8B , or a variant thereof, and wherein said reverse primer is selected from the Jγ primers shown in  FIG. 8B , or a variant thereof. 
     
     
         9 - 13 . (canceled) 
     
     
         14 . A set of nucleic amplification primers capable of amplifying a chromosomal translocation t(14;18)(BCL2-IGH), comprising a forward primer and a reverse primer, wherein said forward primer is selected from the MBR primers, the 3′MBR primers and the mer primers shown in  FIG. 11A , or a variant thereof, and wherein said reverse primer is the J H  consensus primer shown in  FIG. 11A , or a variant thereof. 
     
     
         15 - 19 . (canceled) 
     
     
         20 . A nucleic acid amplification assay, preferably a PCR assay, more preferably a multiplex PCR assay, using at least one set of primers according to  claim 14 . 
     
     
         21 - 27 . (canceled) 
     
     
         28 . A method for detecting a Vγ-Jγ TCRG rearrangement, comprising using one or more sets of primers according to  claim 8  in a nucleic acid amplification assay, preferably a PCR assay, more preferably a multiplex PCR assay. 
     
     
         29 - 33 . (canceled) 
     
     
         34 . A method for detecting a chromosomal translocation t(14;18)(BCL2-IGH), comprising using one or more sets of primers according to  claim 14  in a nucleic acid amplification assay, preferably a PCR assay, more preferably a multiplex PCR assay. 
     
     
         35 - 42 . (canceled) 
     
     
         43 . A nucleic acid amplification assay, preferably a PCR assay, more preferably a multiplex PCR assay, using at least one set of primers according to  claim 8 .