Unit

UNIT 13.7 Detection of Gene Fusions in Acute Leukemia Using Bead Microarrays

  1. German Pihan

Published Online: 1 FEB 2006

DOI: 10.1002/0471142956.cy1307s35

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Pihan, G. 2006. Detection of Gene Fusions in Acute Leukemia Using Bead Microarrays. Current Protocols in Cytometry. 35:13.7:13.7.1–13.7.13.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 FEB 2006
  2. Published Print: JAN 2006

Abstract

This unit describes a method for the rapid and simultaneous detection of hybrid mRNAs (hRNA; also known as gene fusions, RNA fusions, or chimeric RNA), resulting from recurrent chromosome translocations or inversions in acute leukemia. The principal components of the assay are a multiplex RT-PCR, which simultaneously amplifies any of a number of possible hRNA targets, and a liquid bead microarray (LBMA), which is capable of unambiguously identifying the amplified hRNA. The entire procedure from RNA extraction to display of results can be broken into four steps: (1) conjugated-microsphere and LBMA manufacture and quality control, (2) cDNA synthesis, (3) multiplex PCR, and (4) hybridization of multiplex PCR products to the hRNA-detecting LBMA and assay read-out on a flow cytometer. The assay is designed to detect the most common risk-stratifying translocation in pediatric acute lymphoblastic leukemia, satisfying the demand for inclusion of genetic data in the diagnosis of acute leukemia as predicated by the current WHO classification of hematopoietic and lymphoid neoplasms.

Keywords:

  • hybrid mRNA;
  • gene fusions;
  • bead microarray;
  • leukemia