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Minimal Residual Disease (MRD)

Monitoring minimal residual disease and predicting relapse in APL by quantitating PML-RARα transcripts with a sensitive competitive RT-PCR method

Leukemia volume 15, pages 1060–1065 (2001)Cite this article

Abstract

Qualitative RT-PCR methods used for monitoring minimal residual disease (MRD) in APL patients fail to predict relapse in up to 25% of patients in remission. We report here the development and evaluation of a highly sensitive (10−5 and 10−6 with one round and two rounds of PCR, respectively) competitive RT-PCR method to quantitate the PML-RARα fusion transcripts. PML-RARα transcript's levels were normalised to 105 copies of ABL transcript. Serial BM and PB samples from 16 patients with APL and t(15;17) were examined. Presentation samples from three patients (three BM, one PB) showed levels in the range of 0.7 × 106–3.5 × 106 and 1.2 × 105 molecules in BM and PB samples respectively. Serial quantitation of MRD in both BM and PB samples showed significantly lower levels of PML-RARαtranscripts in remission, although the majority of samples remain positive for the PML-RARα transcripts even those in long-term remission (up to 94 months). Levels of PML-RARα in remission samples were up to 2 × 102 and up to 5.2 × 101 molecules in BM and PB respectively. BM and PB samples taken from two patients 2–4 months before relapse showed significantly higher levels of PML-RARαtranscripts (1.2 × 104 molecules in BM; 3.5 × 102, 1.2 × 102 and 1.2 × 103 in PB). The same samples, when tested with a standard qualitative RT-PCR for the amplification of PML-RARα (with a sensitivity of 10−4) produced negative results. This indicates that the qualitative methods would not have predicted relapse in these patients. Our data show that quantitating PML-RARα transcripts with a sensitive method may provide a superior approach for monitoring MRD in APL and identifying patients at high risk of relapse.

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Acknowledgements

This work was supported by the Manchester Leukas-Aid Research Charity. MM was an LRF Clinical Fellow.

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  1. University Department of Haematology, Molecular Oncology Group, Manchester Royal Infirmary, Manchester, M13 9WL, UK

    K Tobal, H Moore, M Macheta & JA Liu Yin

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Tobal, K., Moore, H., Macheta, M. et al. Monitoring minimal residual disease and predicting relapse in APL by quantitating PML-RARα transcripts with a sensitive competitive RT-PCR method. Leukemia 15, 1060–1065 (2001). https://doi.org/10.1038/sj.leu.2402170

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