Detection of the AML translocation (8;21) by two-color fluorescent in situ hybridization
- Univ. of Milan (Italy)
- MRC Molecular Hematology Unit, Oxford (United Kingdom); and others
In the translocation (8;21)(q22;q22) associated with acute myelogenous leukemia (AML), part of the long arm of chromosome 8 is reciprocally translocated onto chromosome 21. At the molecular level the translocation results in the fusion of the 5{prime} region of the AML1 gene on chromosome 21 and almost the entire CDR gene (also ETO or MTG8) on chromosome 8. To detection the translocation at the single cell level, we used two probes, a cosmid clone containing the first five exons of AML1 and a P1 clone containing the entire CDR gene. Hybridization of the two probes to the distal and proximal sides of the translocation breakpoint was expected to highlight the derivative 8q-chromosome in an interphase cell. To demonstrate the ability to identify the translocation in interphase cells using two-color FISH, these two probes were hybridized simultaneously to a cell line containing the 8;21 translocation, Kasumi-1. Each probe was detected with a different color so that their relationship in the sample could be determined within the same interphase cell. Simultaneous hybridization of the CDR and AML1 probes to interphase Kasumi-1 cells resulted in one orange and one green hybridization signal randomly located in the cell, from the hybridization to the normal 8 and 21 chromosomes, and one orange-green pair of signals from the close hybridization of the two probes to the fusion gene on the derivative 8q-chromosome, indicating the translocation. The translocation was identified by an abnormal pairing of the two differently colored signals in the same interphase cell. This technique allows for the detection of the translocation in all cells, not just those arrested in metaphase, and also permits the analysis of a small number of cells. Therefore, useful information can still be obtained from samples not suited for RT-PCR analysis and conventional cytogenetic techniques.
- OSTI ID:
- 133567
- Report Number(s):
- CONF-941009--
- Journal Information:
- American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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