Background : Acute Myeloid Leukemia ( AML ) is amaligmant clonal disorder of immature hematopoietic cells leukemic blasts may express abilities for maturation to avariable degree which leads to morphologic (RQ- PCR ) for monitoring minmal residual disease ( MRD ) in patients with acute promyelocytic leukemia (ARL) ,and to study the clinical application of RQ-PCR of APL for detection of risk of relapse in different phases of treatment , comparing these data with these yielded by conventional qualitative reverse transcriptase – PCR . PATIENT AND METHODS: twenty one consecutive patients diagnosed as acute myeloid leukemia ( M 3) were included in this prospective study patients with cardiac and respiratory diseases were excluded all patients were subjected to the following full history taking , complete clinical examination , some laboratory tests ( complete blood picture & serum creatin ine & serum alaine aminotransferase and serum bilirubin ) . bon marrow asprite ( BMA) for morphology and imunphenotyping ( PT) and cytogenetic studies as well as real – time Qualitative PCR ( RT – PCR ) for detection of PML- RAR agene in BMA at diagnosis and after consolidation were done Quantilative PCR ( RQ – PCR ) in BMA sample after induction phase and consolidation phase to detect normalized copy number (NCN) of PML-RAR α was also done samples taken after informed consent , bonee marrow (BM ) samples were collected from APL patients into tubes containing EDTA antincoagulant before treatment for RT – PCR ,after the induction therepy and after consolidation therapy for RQ – PCR , All patients received AML M3 protocol in the form of induction phase : ATRA : 45 mg / m2 divided into two doses orally till complete remission or maximally for 90 days , and dounroubicin or doxorubicin : 60 mg / m2 I.V. for 3 days (1course ) Also , consolidation phase protocol was given in two courses of dounrobicin or doxorubicin :60 mg/ m2 IV for 3 days every month for two months according to RQ – PCR results after consolidation phase , patients were divided into two groups , group ( A) : NCN OF PML – RAR α1 or leukocyte methotrexate and intermittent ATRA for to 2 years ) During this period patients were kept on follow up for detection of relapse or remission which is defined as : Hematological remission in the form of normalization of peripheral blood and BMA < 5 % blasts and no promyelocyte in peripheral blood and BMA < 5% blasts Hematological relapse was considered in the form of reappearance of > 5% blasts in BMA or promyelocyte RESULTS : there was no statistical difference between the two groups as regarding age ang gender ,Hb levels Platelets count , TLC count , PB promyelcytes (%) ,BM promyelocyte and serum fibrinogen level , there was statistical difference between the two groups as regardingOS , in group A was (13+17.8) and group B was (7.39+13) (p> 0.05) . there was statistical difference between the two groups as regarding DFR ,in group A it was ( 12+24.00) and group B was (8.014+2). There was no statistical correlation between OS & DFR and hemoglobin levels , platelets count , TL , PB promyelocte mean serum fibrinogen level , NCN 1 and BM promyelocyte . there was statistical correlation between OS & DFR and NCN2 (post consolidation )(p < 0.05) there was no statistical correlation between NCN 1 ( post induction) and NCN2 ( post consolidation ) and hemoglobin level , TLC count , platelets count or PB promyelcyte there was no statistical correlation between NCN1 and NCN2 ,mean serum fibrinogen and BM promyelocyte ( p> 0.05) , CONCLUSIONS : these data suggestes that RT- PCR could be used as acomplementary assay for the RQ – PCR approach , especially within thee subgroup with 1-10 NCN furthermore ,it is important to note that the relatively high specifity of RT – PCR assay is not reason enough to substitute ahighly sensitive standardized and high through – put technology such as RQ- PCR recommendations : we recommend other study on alarger scale to study PML- RAR α transcription for risk stratification of relapse acute promyelocytic leukemic Egyptians .
Clinical applications of PML – RAR α Transcript in acute promyelocytic adult Egyptians .j am sci 2013, 9(2) : 247-255 ) .( ISSN :1545-1003) .
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