MRD Quantification of Leukemic Specific Translocations

Numerous publications have demonstrated the feasibility of the RQ-PCR approach to quantify chimeric transcripts resulting from chromosomal translocations occurring in ALL.47,61-63 Although the principles of RQ-PCR are the same whether DNA or RNA is being analyzed, the RT step represents a major assay variable for accuracy of quantification and sensitivity when RNA is used. In fact, it is necessary to correct variations linked to differences in the RNA amount taken for the reaction or, more importantly, in efficiency (or inhibition) during reverse transcription. For this reason, the number of target gene copies has to be normalized using a ubiquitously and constantly expressed housekeeping gene as a reference (e.g., ABL, B2M, and PBGD).6265 Thus, the number of chimeric transcripts will be expressed according to the number of copies of the reference gene transcripts.

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