ZytoLight^® SPEC ERG/TMPRSS2 TriCheck Probe

The ZytoLight^® SPEC ERG/TMPRSS2 TriCheck Probe (PL92) is a direct-labeled FISH probe for the qualitative detection of rearrangements of the human ERG gene at 21q22.2 and the human TMPRSS2 gene at 21q22.3 in formalin-fixed, paraffin-embedded specimens using 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 ERG/TMPRSS2 TriCheck Probe comprises:

• ZyGreen (excitation 503 nm/emission 528 nm) labeled polynucleotides (~10.0 ng/µl), targeting sequences mapping in 21q22.2* (chr21:40,078,039-40,850,582) distal to the ERG breakpoint region;
• ZyOrange (excitation 547 nm/emission 572 nm) labeled polynucleotides (~4.5 ng/µl), targeting sequences mapping in 21q22.13-q22.2* (chr21:39,171,790-39,733,849) proximal to the ERG breakpoint region;
• ZyBlue (excitation 418 nm/emission 467 nm) labeled polynucleotides (~37.0 ng/µl), targeting sequences mapping in 21q22.3* (chr21:43,301,411-44,195,531) distal to the TMPRSS2 breakpoint 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 the human ERG-TMPRSS2 rearrangements by fluorescence in situ hybridization (FISH)

Using suitable filter sets, the hybridization signals of the probe appear green away from the ERG breakpoint region, orange near the ERG breakpoint region, and blue away from the TMPRSS2 breakpoint region.

• Normal scenario: in normal cells during interphase or cells without rearrangements of the respective gene regions, two orange/green fusion signals and two blue signals near the respective fusion signals are observed.
• Aberrant scenario: one 21q22.2 deletion in the 21q22 locus causing the TMPRSS2-ERG fusion is shown by one separate orange signal co-localized with one blue signal, and the loss of one green signal. An ERG translocation without TMPRSS2 involvement is shown by a separated orange signal and a blue signal co-localized with a separate green signal. A non-ERG translocation affecting TMPRSS2 is shown by a separated blue signal not co-localized with the ERG fusion signal.

Overlapping signals may appear yellow.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany