Detection of BCR/ABL Fusion Gene in Chronic Myeloid Leukaemia Using Reverse Transcriptase Polymerase Chain Reaction

Abstract

Background:Chronic myeloid leukemia (CML) is a clonal myeloproliferative expansion of transformed, primitive hematopoietic progenitor cells. It involves myeloid, monocytic, erythroid, megakaryocytic, B-lymphoid, and occasionally T-lymphoid lineages.

Aim: This research was aimed at detecting the BCR-ABL fusion gene in patients with chronic myeloid leukaemia using reverse transcriptase polymerase chain reaction technique.

Material and Method: Eight adults ( aged 45years and above) blood samples were collected from the clinics of Lautech Teaching Hospital, Osogbo, Osun State, Nigeria and University College Hospital, Ibadan, Oyo State, Nigeria. RNA was extracted and purified from CML blood samples using Axyprep multisource Total RNA miniprep kit, USA procedures were followed.

Result: At the end of the reaction, amplifications were not observed at the annealing temperature of 55 ºC. The only bands were the primer sets and some DNA contaminations at the RNA controls for both multiplex and nested PCRs.

Conclusion: Total RNA extraction from the blood sample is crucial to the success of RT-PCR, without this, the study cannot run to completion. Meanwhile, in this experiment, it was observed that there were no bands when the eluted total RNA was run on gel electrophoresis. This indicates that there were no extraction of RNA. Rapid degradation of RNA is the most important factor impeding the analysis of gene expression in human cells and tissues.