Detection of the hybrid BCR/ABL messenger RNA in single CFU-GM colonies using the polymerase chain reaction.
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.
Leukaemia Research Fund Centre, Institute of Cancer Research, London, UK.
A
A
Hernandez
J
J
Osterholz
C M
CM
Price
L M
LM
Wiedemann
M Y
MY
Gordon
J M
JM
Goldman
G J
GJ
Morgan
D001853:Bone Marrow / Q000473:pathology; D002478:Cells, Cultured; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006098:Granulocytes / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000473:pathology; D008264:Macrophages / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl