Chronic myelogenous leukemia (CML)is a myeloproliferative disorder in which the hematopoetic stem cell has undergone neoplastic transformation. An interesting aspect of CML ia the presence of specific chromosome abnormality called Philadelphia (Ph 1+)chromosome in the leukemia cells of most patients.
A kit based RT-PCR (Promega)system was used in the qualitative detection of BCR-abl mRNA using the following set of primers: Bcr exon bl (NBI+); Abl exon a3 (Abl3); Bcr specific (B2A); ABL specific (CA3) and positive control primers: CA3 and A2, The design of all primers were based according to the predominant hybrid transcripts in CML patients (chimera), namely: b2a2 and b3a2. There were 25 patients included in the study, 8(32%) of which showed positivity in the Bcr-abl translocation in either peripheral blood, bone marrow or for both specimens while 16 specimens showed no translocation. The rest of the samples did not yield any RNA due to probable technical difficulties. The results of thid study suggest a potential use of peripheral blood specimen for RT-PCR in the detection of BCR/abl translocation especially in cases when bone marrow aspiration is a dry tap. Problems on undetected fusion proteins in CML in this study like ela2 and el19a2 may be addressed using different array of primers.
The general objective of this study is to determine the % positivity of BCR-ABL gene in the peripheral blood and bone marrow aspirates among clinically diagnosed CML patients.The specific objectives are to isolate total RNA from peripheral blood specimens from normal and CML patients: to determine qualitatively the presence of BCR-ABL mRNA using RT-PCR (Access system by PROMEGA): and to compare the % positivity of BCR-ABL mRNA in bone marrow aspirates and peripheral blood in clinically diagnosed CML patients.
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