Competitive PCR method for residual disease quantification.

A competitor was constructed by PCR using genomic DNA from the malignant clone as template together with a clone-specific primer and primer-XhoI to introduce a restriction site. The first amplification in the seminested PCR was done with the clone-specific primer and primer-1 and the second amplification with the clone-specific primer and primer-2. The PCR products from the competitor are cleaved by the restriction enzyme XhoI; the PCR products from the malignant clone remain uncleaved. The MRD level can be determined from the lanes with equal amplification of the competitor and malignant clone.14