RT-PCR method with increased sensitivity shows persistence of PML-RARA fusion transcripts in patients in long-term remission of APL.
RT-PCR methods have been developed, to date, by various groups to amplify the PML-RARA fusion gene produced by the t(15;17) in APL patients. However, these methods lack the necessary sensitivity to detect minimal residual disease (MRD) below the level of 1 leukaemic cell in 10(4) cells. Patients who test positive by these methods after treatment are likely to relapse. However, up to 25% of patients who test negative after treatment relapse within a short period. We have developed a 'hot-start' RT-PCR method for the amplification of PML-RARA with increased sensitivity at the level of two leukaemic cells in 10(6) cells. Using this method we were able to detect MRD in seven out of 15 patients tested in remission. Of the 11 patients in medium to long-term remission, five patients tested positive. None of these 11 patients tested positive with the standard RT-PCR. These results show that some patients in remission of APL continue to express PML-RARA even in long-term remission, when they can be considered clinically 'cured' of their disease.
Tobal
K
K
University Department of Haematology, Manchester Royal Infirmary, UK.
Liu Yin
J A
JA
D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics*; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics; D018365:Neoplasm, Residual; D015514:Oncogene Proteins, Fusion / Q000032:analysis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D009363:Neoplasm Proteins; D015514:Oncogene Proteins, Fusion; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein