RRML - Real-time qPCR for assessment of minimal residual disease in acute myeloid and lymphoid leukemia

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Nr. 19(4)/2011

Real-time qPCR for assessment of minimal residual disease in acute myeloid and lymphoid leukemia

Dumitru Jardan, Rodica Talmaci, Cerasela Jardan, Daniel Coriu, Anca Colita, Andrei Colita, Constantin Arion

Abstract:

Evaluation of minimal residual disease is of major importance in the course of acute leukemia treatment. There are a number of technologies used in this regard, all of them exhibiting different limitations. Real time qPCR is one of the main technologies enabling gene expression analysis and is well suited for minimal residual disease monitoring of acute leukemia patients. cDNA of eight most common fusion genes was cloned - PML-RARα, TEL-AML1, AML-ETO, E2A-PBX1, SIL-TAL1, CBFβ-MYH11, MLL-AF4, BCR-ABL1. Serial dilution of cloned plasmids were prepared and used as standards for real-time qPCR. Sensitivity and overall performance of the method was evaluated as previously described. Overall testing demonstrated robustness of TaqMan technology for gene expression analysis. High reproducibility with low levels of both inter and intra run variation was obtained. Sensitivity exhibited by the assay was comparable to nested PCR which allowed early relapse detection for a number of patients.

Keywords: real-time qPCR,acute leukemia,minimal residual disease.

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Jardan D, Talmaci R, Jardan C, Coriu D, Colita A, Colita A, et al. Real-time qPCR for assessment of minimal residual disease in acute myeloid and lymphoid leukemia. Rev Romana Med Lab. 2011;19(4):349-57