ZytoLight^® SPEC EGFR/CEN 7 Dual Color Probe

The ZytoLight^® SPEC EGFR/CEN 7 Dual Color Probe (PL15) is a direct-labeled FISH probe designed for the qualitative detection of amplifications involving the human EGFR gene and chromosome 7 alpha satellites in formalin-fixed, paraffin-embedded specimens by fluorescence in situ hybridization (FISH). The 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 EGFR/CEN 7 Dual Color Probe comprises:

• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~10 ng/µl), targeting sequences mapping in 7p11.2* (chr7:54,912,555-55,548,375) harboring the EGFR gene region;
• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~1.5 ng/µl), targeting sequences mapping in 7p11.1-q11.1 specific for the alpha satellite centromeric region D7Z1 of chromosome 7;
• 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 amplifications involving the human EGFR gene and chromosome 7 alpha satellites, via FISH.

Using suitable filter sets, the hybridization signals of the probe appear green at the EGFR gene region and orange at CEN 7.

• Normal scenario: in normal cells during interphase or cells without an amplification of the EGFR gene region, two green and two orange signals are observed.
• Aberrant scenario: in cells with an amplification of the EGFR gene region, multiple copies of the green signal or green signal clusters are observed.

Overlapping signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany