Method of HER2 gene copy number quantification in samples with indeterminate ISH

Current oncology is focused towards the search and use of predictive biomarkers that could determine suitable patients for targeted therapy. One of the well-known predictive biomarker is HER2 gene, localized on 17q, which is amplified in 15-20% breast cancer patients. HER2 amplification is poor prognostic factor but predictor of good response to anti-HER2 therapy (trastuzumab, pertuzumab and lapatinib) which significantly increases survival rates in both palliative and adjuvant settings. Anti-HER2 agents are approved for treatment of HER2- positive breast cancer patients usually determined by immunohistochemistry and in situ hybridization (ISH). The proper determination of HER2 status is therefore fundamentally important.

The HER2 DNA quantification kit was developed as a complementary test method for the quantification of HER2 gene in breast cancer patient samples which cannot be reliably evaluated by ISH. The kit works on the basis of the three duplex quantitative real-time polymerase chain reactions (qPCR) and is applicable for DNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples. The HER2 gene copy number status is compared with three reference genes – GCS1 (chromosome 2), DCK (chromosome 4) and EPN2 (chromosome 17). The kit reliably detects HER2 gene amplification in samples containing at least 5% of strongly positive cells (approximately 20 HER2 gene copies per cell). High sensitivity and specificity levels were validated using 223 breast cancer patient samples.

The HER2 DNA quantification kit was developed as a complementary test method for the quantification of HER2 gene in breast cancer patient samples which cannot be reliably evaluated by ISH. The kit works on the basis of the three duplex quantitative real-time polymerase chain reactions (qPCR) and is applicable for DNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples. The HER2 gene copy number status is compared with three reference genes – GCS1 (chromosome 2), DCK (chromosome 4) and EPN2 (chromosome 17). The kit reliably detects HER2 gene amplification in samples containing at least 5% of strongly positive cells (approximately 20 HER2 gene copies per cell). High sensitivity and specificity levels were validated using 223 breast cancer patient samples.

CZ 28596

Laboratory scale, validation study on patient samples.