real-time quantitative PCR
Abstract
The purpose of this paper is to enlighten us on how real-time quantitative PCR can be useful in the detection, assessment and diagnosis of chronic myeloid leukemia. Chronic myeloid leukemia was the first neoplasm to be associated with a single specific acquired genetic lesion. It is one of the well understood myleproliferative disorders at the molecular level. The disease originates from the transformation of a hematopoietic stem cell with consequent expanding myelopoiesis. Molecular monitoring of BCR- ABL is becoming common in the diagnosis and assessment of treatment responses of patients suffering from chronic myeloid leukemia (CML). This practice has become relevant especially when the residual levels of fall below the level of detection by cytogenetic analysis. Forty-two CML patients consisting up of eighteen males (42.86%) and twenty four females (57.14%) all aged between 7-75 years were selected for the study and placed under observation for the response to imatinib treatment. The patients were subjected to Multiplex RT-PCR (reverse-transcriptase PCR) and they were all found to contain either e13a2 or the e14a2, which could be analyzed by a single Taqman probe based quantitation kit (Geno-Sen’s) to quantitate the BCR-ABL transcript load. The Multiplex RT-PCR and peripheral blood cytogenetics providing specific and sensitive detection of BCR-ABL fusion transcripts and metaphase signal load respectively were used as parallel reference tools to verify the q-PCR findings. There was 100% concordance between the multiplex RT-PCR and the q-PCR as every positive RT-PCR assay for a transcript reflected as q-PCR load of above 0% for that transcript. q-PCR also managed to show a strong Pearson correlation with the cytogenetic response.