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Plasma RNA as an alternative to cells for monitoring molecular response in patients with chronic myeloid leukemia

From the Hematopathology Department, Quest Diagnostics Nichols Institute, San Juan Capistrano, CA, USA (WM, RT, MG, IJ, MA); Leukemia Department, M.D. Anderson Cancer Center, University of Texas, Houston, TX, USA (MK, HK, JC, SOB, FG)

Correspondence: Maher Albitar, MD, Quest Diagnostics Nichols Institute, 33608 Ortega Highway, Rm#108B, San Juan Capistrano, CA 92690-6130 USA. E-mail: maher.x.albitar{at}questdiagnostics.com

Background and Objectives: : Quantitation of BCR-ABL mRNA is emerging as the standard of care to monitor the status of chronic myeloid leukemia (CML). Peripheral blood plasma was analyzed in this study because of previous detection of nucleic acids and proteins from tumor cells in plasma samples.

Design and Methods: : Reverse transcriptase polymerase chain reaction was used to establish ratios of BCR-ABL:ABL mRNA in peripheral blood cells and plasma, and absolute levels of BCR-ABL mRNA per unit volume of plasma. Samples from 160 CML patients and 180 control individuals without CML were tested. Cells and plasma samples from 93 of the CML patients were re-analyzed 3–12 months after imatinib treatment.

Results: : Ratios of BCR-ABL:ABL mRNA in paired cell and plasma samples of the 160 CML patients correlated significantly (r=0.83; p<0.001). When results were compared directly using the sign test, the pre-therapy plasma results were significantly different from those from peripheral blood cells (p=0.028), but not bone marrow cells (p=0.119). Absolute levels of BCR-ABL mRNA in plasma strongly correlated with many laboratory characteristics in pre-therapy CML patients. Higher BCR-ABL: ABL ratios were detected in plasma samples at all time points after treatment, although this was significant only at 3 months (p=0.0003). In cases in which results from the assays disagreed, minimal residual disease was detected in plasma samples significantly more frequently than in cell samples (p<0.001).

Interpretation and Conclusions: : Plasma was a reliable source for monitoring BCR-ABL mRNA levels. Minimal residual disease detection from plasma was more sensitive than from cell samples. Our results suggest that absolute levels of BCR-ABL mRNA per unit volume of plasma may reflect tumor load.

Key words: BCR-ABL, RT-PCR, chronic myeloid leukemia, imatinib, plasma.

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