Detection of the BCR/ABL fusion gene in chronic myeloid leukemia by RNA polymerase chain reaction

Summary

The bcr/abl fusion gene in 20 patients with chronic myeloid leukemia (CML) was detected by RNA polymerase chain reaction, which used mRNA as the starting material to generate cDNA with reverse transcriptase followed by PCR amplification (RNA/PCR). Amplification of a sequence spanning the bcr/abl junction region was achieved by using peripheral blood cells as the sourse of mRNA from all 20 patients with CML, including 3 cases of Ph(-) CML, and cell line K562 was derived from patients with CML. No amplification was seen when mononuclear cells from 3 normal individuals, 2 patients with lymphoma and cell line HL-60 were used. The presence or absence of bcr exon 3 in the fusion mRNA was determined by the size of the amplified fragments. Of the 20 CML patients, 15 showed only the 165-bp amplified band (indicating retention of bcr exon 3), done showed only the 90-bp amplified band, and A showed both 165-bp and 90-bp bands. Both bands Were seen more frequently in blast crisis than in remission and chronic phase.