Product description

The ESR1 and PIK3CA mutations have been carefully selected to cover the most clinically relevant variants. With this control material, scientists and diagnosticians can improve the sensitivity and specificity of their detection methods while ensuring consistent performance.

The material can be diluted with wild-type DNA (wt) as needed to achieve the desired allele frequency. The corresponding wild-type DNA is already included in the set. Additionally, the material is ideal for use in minimal residual disease (MRD).

1. Combination of Mutations: By combining ESR1 and PIK3CA mutations in a single product, efficient and time-saving quality control is enabled.
2. Flexibility in Application: The material has been specifically optimized for highly sensitive methods such as NGS and digital PCR, ensuring precise results with minimal background noise.
3. Optimal Reproducibility: Manufacturing follows strict quality guidelines to ensure consistent performance and minimal batch-to-batch variability.
4. Applications: Perfectly suited for clinical studies, research, validation, and routine diagnostics.

This product includes the following set of separate vials:

ESR1 Mutations:

Mutations in the ESR1 gene are recognized biomarkers for endocrine resistance in hormone receptor-positive breast cancer. They frequently occur after therapy with aromatase inhibitors and play a central role in monitoring therapy response and resistance development.

PIK3CA Mutations

PIK3CA mutations are among the most common genetic alterations in breast cancer and other solid tumors. These mutations are predictive markers for the efficacy of PI3K inhibitors and are becoming increasingly significant in personalized medicine.

The combination of both mutations in a single control material allows for comprehensive evaluation and optimization of molecular diagnostic tests.

Buffer: Tris-EDTA (10 mM Tris, 1 mM EDTA), pH 8.0

Storage: 2–8 °C

Shelf Life: Stable up to 24 months from the date of manufacture (as supplied)

Quality Assurance

• Concentration: 10.0 ng/μl
• DNA quantity per vial with mutated DNA: 250 ng
• DNA quantity per vial with wild-type DNA: 1250 ng
• Fragment size determination: TapeStation electrophoresis (Agilent)
• Allele frequency/copy number: dPCR (metrologically recognized primary reference measurement method) (3)
• Quantification (metrologically traceable) via fluorometric dsDNA measurement (Qubit)

Technical Background

Wild-type cfDNA for the specified ESR1 and PIK3CA mutations (GRCh38).

Use as a Complete Workflow Control Material

For full workflow controls, we recommend using our human DNA-free plasma products. Mix the DNA with DNA-free plasma as needed to produce plasma materials with the required DNA concentrations.

Link to DNA-Free Plasma Products

OEM Options

If you need materials as in-kit controls or as third-party controls for the validation of your kits in customer laboratories, please contact us at: sales@sens-id.com

For Use with LDTs for:

1. Evaluation of test accuracy
2. Verification of analytical sensitivity and specificity
3. Monitoring reproducibility
4. Validation of new batches and reagents
5. Ensure quality assurance
6. Error analysis and troubleshooting
7. When developing and performing LDTs for identifying therapeutically relevant mutations in DNA, the use of QCMs is especially important to ensure result reliability and patient safety.

For Use with CE-IVD-Labeled Assays

CE-IVD (Conformité Européenne – In Vitro Diagnostics) labeled assays are commercially manufactured in vitro diagnostic assays that have received European conformity certification. The use of an external quality control material, such as that provided by SensID, alongside CE-IVD labeled assays in a laboratory offers several advantages:

1. Independent Evaluation: External QCMs are independently manufactured from the CE-IVD test manufacturer. This independence ensures the QCMs are unbiased and not influenced by the assay manufacturer, enabling an objective evaluation of laboratory performance.
2. Validation of Manufacturer Claims: External QCMs can be used to independently verify manufacturer claims regarding assay performance, such as accuracy, precision, sensitivity, and specificity.
3. Continuous Improvement: External QCMs provide laboratories with a tool for continuous monitoring and improvement of their assay processes. If discrepancies or issues are identified using external QCMs, laboratories can take corrective actions to review and improve assay performance.
4. Regulatory Compliance: In some regions, regulators recommend or require the use of external QCMs as part of the quality control process. Adhering to such recommendations or requirements ensures the laboratory meets regional quality and accreditation standards.
5. Risk Mitigation: By using external QCMs, laboratories reduce the risk of relying solely on QCMs provided by the manufacturer, which may not cover all potential sources of variability or adequately verify assay performance across the entire analytical range. For example, plasmids used as positive and negative controls in kits do not reflect the complexity of a patient sample, including the full genomic background. When in doubt, ask your assay provider about the control materials supplied.

Confidence in Results: Using external QCMs contributes to overall confidence in the accuracy and reliability of test results obtained with CE-IVD assays. This confidence is critical when these tests are used for patient diagnosis and treatment decisions.

Additional Information

In several retrospective cohorts, mutations in estrogen receptor 1 (ESR1) were identified in approximately 11%-55% of patients with estrogen receptor (ER)-positive metastatic breast cancer treated with aromatase inhibitors (AI) (1).

According to the results of the randomized Phase III EMERALD trial, progression-free survival (PFS) was prolonged in all patients (Hazard Ratio = 0.70; 95% CI, 0.55 to 0.88; P = .002) and in patients with ESR1 mutations (Hazard Ratio = 0.55; 95% CI, 0.39 to 0.77; P = .0005). Elacestrant is the first oral selective ER degrader to demonstrate significant PFS improvement compared to standard of care (SOC) in both the overall population and in patients with ESR1 mutations with manageable safety in a Phase III trial of ER-positive/HER2-negative advanced breast cancer (2).

PIK3CA mutations occur in approximately 40% of patients with hormone receptor (HR)-positive, HER2-negative breast cancer, with mutations primarily located in exons 9 and 20. PIK3CA mutations are also prevalent in other tumor entities, including endometrium, colon, urinary tract, cervix, and uterus (4–8).

References

1. Clinical Implications of Monitoring ESR1 Mutations by Circulating Tumor DNA in Estrogen Receptor Positive Metastatic Breast Cancer: A Pilot Study
2. Elacestrant (oral selective estrogen receptor degrader) Versus Standard Endocrine Therapy for Estrogen Receptor-Positive, Human Epidermal Growth Factor Receptor 2-Negative Advanced Breast Cancer: Results From the Randomized Phase III EMERALD Trial
3. Assessment of Digital PCR as a Primary Reference Measurement Procedure to Support Advances in Precision Medicine
4. The Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature. 2012;490(7418):61-70.
5. Tolaney S, Toi M, Neven P, et al. Presented at: 2019 American Association for Cancer Research (AACR) Annual Meeting; March 29 – April 3, 2019; Atlanta, GA.
6. Di Leo A, Johnston S, Seok Lee K, et al. Buparlisib plus fulvestrant in postmenopausal women with hormone-receptor-positive, HER2-negative, advanced breast cancer progressing on or after mTOR inhibition (BELLE-3): a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Oncol. 2018;19(1):87-100.
7. Moynahan ME, Chen D, He W, et al. Correlation between PIK3CA mutations in cell-free DNA and everolimus efficacy in HR+, HER2− advanced breast cancer: results from BOLERO-2. Br J Cancer. 2017;116(6):726-730.
8. Ebubekir Dirican, Mustafa Akkiprik & Ayşe Özer. Mutation distributions and clinical correlations of PIK3CA gene mutations in breast cancer. Tumor Biol. 2016. Volume 37, pages 7033–7045