Abstract
Nested reverse-transcriptase polymerase chain reaction (rt-PCR) was performed on 58 leukemia patients at BIRDEM Laboratory, as a pioneering work in Bangladesh. Thirty of themwere examined for the presence of BCR-ABL being clinically and morphologically diagnosed as chronic myeloid leukemia (CML) and 28 for PML-RARalpha fusion transcripts being clinically and morphologically diagnosed as acute promyelocytic leukemia (APL/ AML M3). The cases were selected for targeted therapy with imatinib mesylate and all-Trans retinoic acid (ATRA) to treat CML and APL respectively. Samples were received either before commencement or during therapy. In the positive cases, amplified DNA products were visible after gel electrophoresis and were reported accordingly. In case of BCR-ABL, positive results were found for five out of six (83.33%) untreated cases and 11 out of 24 (45.83%) treated cases. Positive results for PML-RARalpha were found for 12 out of 14 (85.70%) untreated cases and 11 out of 16 (68.75%) treated cases. A strong positive correlation was found between duration of treatment and negativity of PCR results in both the cases. In present times, the detection of minimal residual disease in patients undergoing treatment for hematological malignancies has become an important goal, not only to monitor the effectiveness of therapy but also to detect an impending relapse. This is the first time in Bangladesh that rt-PCR method is being employed to detect or monitor the presence of abnormal fusion genes in hematological malignancies.
Highlights
Diagnosis of hematological malignancies is not complete unless morphological examination is supplemented by further studies like immunophenotyping, karyotyping and/or molecular studies, etc
RNA isolated from bone marrow or peripheral blood samples of 26 patients were examined for the presence of the breakpoint cluster regions (BCR)-ABL and 24 for PML-RARα fusion transcript
Group I: 14 patients belonged to this group, who had been provisionally diagnosed as acute promyelocytic leukemia (APL) and the samples collected prior to initiation of therapy
Summary
Diagnosis of hematological malignancies is not complete unless morphological examination is supplemented by further studies like immunophenotyping, karyotyping (conventional cytogenetics) and/or molecular studies, etc. Chronic myeloid leukemia (CML) and acute promyelocytic leukemia (APL) are two prototypes of such diseases Molecular methods such as fluorescence in situ hybridization (FISH), reverse-transcriptase polymerase chain reaction (rt-PCR), and real-time quantitative rt-PCR have been used to detect the chimeric genes in question or their transcripts. Among the various cytogenetic and molecular tests available for diagnosis and monitoring of hematological malignancies, polymerase chain reaction (PCR) based amplification assays are the most sensitive of all laboratory approaches, followed by flow cytometry, FISH, and cytogenetics, with associated detection thresholds of about one in one million, one in several thousand, one in several hundred, and one in 20, respectively[1]. As the results may vary largely depending on expertise, manual errors are more likely
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