Real‐time reverse transcription polymerase chain reaction detection and quantification of t(1;19) (E2A–PBX1) fusion genes associated with leukaemia

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Abstract
A real‐time reverse transcription polymerase chain reaction (RT‐PCR) method is described that enabled the detection and quantification of E2A–PBX1 fusion gene transcripts associated with t(1;19). The method was highly reproducible and offered exceptional sensitivity at 5 fg of fusion transcript per reaction, without the need for a nested PCR primer design. To illustrate the usefulness of this new technology the E2A–PBX1 fusion gene transcript expression level for several human leukaemia cell lines that are positive and negative for cytogenetically detectable t(1;19) was determined. The RCH–ACV had a threefold higher expression of E2A–PBX1 transcripts (600 transcripts per cell) than the other t(1;19) positive 697 (150 transcripts per cell). The only other cell line with detectable E2A–PBX1 was CEM, but the level of expression was < 1 transcript per cell.