تشخيص كمي رونويس هاي ژن bcr-abl اصلي در مبتلايان به لوسمي ميلوئيد مزمن از طريق competitive RT-PCR

Quantitative detection of Major BCR-ABL gene transcripts by competitive RT-PCR in chronic myeloid leukemia (CML) patients

Chronic Myeloid Leukemia (CML) is produced by the chromosomal translocation and fusion of bcr/abl genes. The purpose of this study was the development of a simple and low-cost technique for quantitative detection of bcr/abl transcripts in CML patients. This study, we used the competitive RT-PCR technique to determine the number of bcr/abl transcripts. The internal control was designed from a non-human DNA source with the same flanking sequences but a smaller size than the target segment. After amplification and purification of internal control DNA, PCR products were quantified based on copy numbers. In order to optimize the reaction condition, competitive RT-PCRs were carried out separately on target and internal control DNAs each with specific copy numbers, and final products of all concentrations were run on agarose gel electrophoresis. The simultaneous reactions of target RNA with different copy numbers of internal control DNA showed that the copy numbers of target RNA can be determined by comparison of dye density emission of DNA bands(target and control DNA) on the gel. This study shows that competitive RT-PCR is a partially efficient and cheap method for quantitative determination of bcr/abl transcripts, compared to other methods such as Real-time PCR.