Real-time reverse transcription polymerase chain reaction detection and quantification of t(1;19) (E2A-PBX1) fusion genes associated with leukaemia.
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.
Curry
J D
JD
Division of Environmental Health Sciences, School of Public Health, University of California, Berkeley, CA 94720-7360, USA.
Glaser
M C
MC
Smith
M T
MT
D002460:Cell Line; D002878:Chromosomes, Human, Pair 1; D002888:Chromosomes, Human, Pair 19; D003199:Computer Systems; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D007938:Leukemia / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic
D018398:Homeodomain Proteins; D015514:Oncogene Proteins, Fusion; C089847:E2A-Pbx1 fusion protein