Detection of the hybrid BCR/ABL messenger RNA in single CFU-GM colonies using the polymerase chain reaction

Exp Hematol. 1990 Nov;18(10):1142-4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Marrow / pathology
  • Cells, Cultured
  • Fusion Proteins, bcr-abl / genetics*
  • Granulocytes / metabolism
  • Hematopoietic Stem Cells / chemistry*
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / pathology
  • Macrophages / metabolism
  • Polymerase Chain Reaction*
  • RNA, Messenger / analysis*

Substances

  • RNA, Messenger
  • Fusion Proteins, bcr-abl