ZytoLight^® SPEC BCR/ABL1 Dual Color Dual Fusion Probe

The ZytoLight^® SPEC BCR/ABL1 Dual Color Dual Fusion Probe is a direct-labeled fluorescence in situ hybridization (FISH) probe designed for the qualitative detection of the t(9;22)(q34.1;q11.2) chromosomal translocation involving the BCR and ABL1 genes, commonly associated with chronic myeloid leukemia (CML). For optimal performance, the probe is intended for use with the ZytoLight^® FISH-Cytology Implementation Kit.


ZytoLight^® probes enable the precise identification of genetic abnormalities—including translocations, deletions, amplifications, and chromosomal aneuploidies—in formalin-fixed, paraffin-embedded (FFPE) tissue sections or cytology specimens, without the need for additional antibody-based detection.

Components

The ZytoLight SPEC BCR/ABL1 Dual Color Dual Fusion Probe comprises:

• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~12 ng/µl), targeting sequences mapping in 22q11.22-q11.23* (chr22:23,000,029-24,431,064) harboring the BCR gene region;
• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~6 ng/µl), targeting sequences mapping in 9q34.11-q34.13* (chr9:133,223,081-134,103,849) harboring the ABL1 gene region;
• Formamide based hybridization buffer

*according to Human Genome Assembly GRCh37/hg19

Principle

The fluorescence in situ hybridization (FISH) method enables the identification and visualization of particular nucleic acid sequences within cell samples. DNA fragments labeled with fluorescent dyes, known as FISH probes, are co-denatured with their complementary target DNA strands in the sample and then allowed to hybridize. Following hybridization, non-specific and unbound probe fragments are removed through stringency washes. The DNA is then counterstained with DAPI, and the hybridized probes are detected using a fluorescence microscope equipped with filters matched to the specific fluorochromes used to label the FISH probes.

For the qualitative detection of translocation t(9;22)(q34.1;q11.2) by fluorescence in situ hybridization (FISH)

Using suitable filter sets, the probe’s hybridization signals display as green for the BCR gene region and orange for the ABL1 gene region.
Normal scenario: normal cells or in cells lacking a translocation involving these gene regions, two distinct signals are observed: one green and one orange during interphase.
Aberrant scenario: where gene fusion has occurred, the signal pattern consists of one green signal, one orange signal, and two combined orange/green fusion signals.
Overlapping orange and green signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany