A rapid RT-PCR based method for the detection of BCR-ABL translocation.
OBJECTIVE
To optimise a one step reverse transcriptase polymerase chain reaction (RT-PCR) protocol for BCR-ABL chimaera detection.


METHODS
Compared with published RT-PCR procedures, this novel approach has at least two advantages. First, the same enzyme is used for both reverse transcription and PCR. Second, amplification of the target (BCR-ABL chimaera) and control gene (ABL) is performed simultaneously in the same tube.


RESULTS
On testing 40 chronic myelogenous leukaemia patients and 10 healthy donors there was a specificity for the newly developed technique. In addition, dilution experiments demonstrated that the protocol was highly sensitive.


CONCLUSIONS
The suggested one step PCR strategy is a simple and reliable way to reveal BCR-ABL chimaeras.
Sidorova JYu
Institute of Haematology and Transfusiology, St Petersburg, Russia.
Saltykova
L B
LB
Lyschov
A A
AA
Zaritskey AYu
Abdulkadyrov
K M
KM
Blinov
M N
MN
D012321:DNA-Directed RNA Polymerases; D016313:Genes, abl; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D015513:Oncogene Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D051562:Proto-Oncogene Proteins c-bcr; D012680:Sensitivity and Specificity; D013997:Time Factors; D014178:Translocation, Genetic
D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr; D012321:DNA-Directed RNA Polymerases