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Applicability of an absolute quantitative procedure to monitor intra‐individual bcr/abl transcript kinetics in clinical samples from chronic myelogenous leukemia patients
Author(s) -
Kreuzer KarlAnton,
Lass Ulrich,
Nagel Stefan,
Ellerbrok Heinz,
Pauli Georg,
PawlaczykPeter Bärbel,
Siegert Wolfgang,
Huhn Dieter,
Schmidt Christian Andreas
Publication year - 2000
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(20000601)86:5<741::aid-ijc22>3.0.co;2-1
Subject(s) - taqman , chronic myelogenous leukemia , abl , breakpoint cluster region , real time polymerase chain reaction , fusion protein , complementary dna , biology , leukemia , cancer research , minimal residual disease , immunology , genetics , gene , tyrosine kinase , recombinant dna , receptor
Chimeric bcr/abl fusion proteins are thought to be the molecular 'pathogen' of chronic myelogenous leukaemia (CML). Expression levels of the respective fusion RNAs reflect disease progression as well as remission upon therapeutic intervention in CML patients. However, there is no quick and reliable method that would allow the quantitative routine monitoring of bcr/abl hybrid transcripts. A fluorescent probe‐based PCR assay (TaqMan) has been described to quantitfy the exact amount of target sequences. We have established TaqMan real‐time RT‐PCRs for M‐bcr/abl (b2a2, b2a3, b3a2, b3a3) and m‐bcr/abl (e1a2) fusion transcripts as well as for β‐actin. All PCRs quantified as little as 10 copies/100 ng total cDNA. In order to investigate whether this procedure is appropriate for routine diagnostic monitoring, we performed retrospective measurements on 9 documented CML disease courses. Our data show that ongoing or relapsing CML is paralleled by increasing peripheral levels of bcr/abl fusion RNAs. Furthermore, sucessful anti‐leukemic treatment is reflected by decreasing absolute bcr/abl transcript numbers. In contrast with conventional bcr/abl PCR techniques we could distinguish single positive values and gradually increasing copy numbers. Int. J. Cancer 86:741–746, 2000. © 2000 Wiley‐Liss, Inc.