Detection of the hybrid BCR/ABL messenger RNA in single CFU-GM colonies using the polymerase chain reaction
Hernandez, A.; Osterholz, J.; Price, C.M.; Wiedemann, L.M.; Gordon, M.Y.; Goldman, J.M.; Morgan, G.J.
Experimental Hematology 18(10): 1142-1144
1990
ISSN/ISBN: 0301-472X PMID: 2209769 Document Number: 357715
In order to study which hemopoietic precursor cells express the hybrid BCR/ABL fusion mRNA we have developed a technique based on the polymerase chain reaction (PCR) for the examination of single hemopoietic colonies grown on semi-solid agar. The technique was developed by examining single CFU-GM colonies grown from newly diagnosed patients with chronic myeloid leukaemia (CML). RNA was isolated from individual 14 day colonies and reverse transcribed to a complementary DNA (cDNA) copy which formed the substrate for a PCR. We have studied 3 cases of CML using this method and have found that 5 out of 5, 9 out of 10 and 20 out of 23 colonies examined were positive. Thus we describe a simple and useful technique for the study of gene expression in a limited number of hemopoietic precursor cells.