Control genes for MRD in leukemia

Whether absolute or relative quantitation is carried out, the choice of control gene is very important. Older publications refer to sequences such as P-actin, GAPDH or 18S ribosomal RNA for gene expression assays. The first two have pseudogenes and inaccuracies may result from DNA contamination, the latter is expressed at very high levels and is not a suitable control for single copy genes without limiting primer concentrations. Both test and control genes must amplify with the same efficiency for the AACt method to be valid. Another important consideration is that the expression of the control gene may be influenced by the treatment the patient is

BCRABL timecourse

1000

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Figure 16.2

BCRABL/ABL Log reduction

BCRABL/ABL Log reduction

0.001

Time (months)

Tracking response to therapy in a patient with CML.

undergoing and should remain stable during the course of treatment. The Europe Against Cancer (EAC) project addressed the choice of control genes (Beillard et al, 2003); ABL, GUS, and B2M (p-2- microglobulin) emerged as the most suitable. For genomic DNA targets recommended control genes include B2M, ALB (albumin), and HBB (p-globin).

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