ZytoLight^® SPEC BCL2/IGH Dual Color Dual Fusion Probe

The ZytoLight^® SPEC BCL2/IGH Dual Color Dual Fusion Probe is a direct-labeled fluorescence in situ hybridization (FISH) probe designed for the qualitative detection of the t(14;18)(q32.3;q21.3) translocation involving the IGH and BCL2 genes. This probe enables the identification of this specific genetic aberration in formalin-fixed, paraffin-embedded (FFPE) tissue samples. For optimal performance, this probe is intended for use with the ZytoLight^® FISH-Tissue 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 BCL2/IGH Dual Color Dual Fusion Probe comprises:

• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~6 ng/µl), targeting sequences mapping in 18q21.33* (chr18:60,180,078-61,507,691) harboring the BCL2 gene region;
• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~12 ng/µl), targeting sequences mapping in 14q32.33* (chr14:105,462,169-106,995,000) harboring the IGH locus;
• 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.

This probe is intended for use in formalin-fixed, paraffin-embedded specimens to qualitatively detect translocation t(14;18)(q32.3;q21.3) involving the human IGH and BCL2 genes, via FISH.

Using suitable filter sets, the probe’s hybridization signals are seen as orange for the BCL2 gene region and green for the IGH locus.

• Normal scenario: In interphase nuclei of cells without translocations affecting these gene regions, two distinct green signals and two distinct orange signals are observed.
• Aberrant scenario: The presence of a gene fusion is shown by one separate orange signal, one separate green signal, and two orange/green fusion signals.

Overlapping orange and green signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany