Real-time clinical validation of the Geneverify blood mRNA test for breast cancer screening and diagnosis

Background: Early detection remains the cornerstone of successful breast cancer management. We conducted a real-time clinical validation of the plasma cell-free mRNA-based Geneverify test for early screening and detection of breast cancer. This next-generation approach offers rapid, precise, and non-invasive results, potentially transforming the current diagnostic landscape by reducing dependence on surgical biopsies and enhancing early diagnosis. Methods: We evaluated the real-time expression of 30 different breast cancer associated genes in plasma samples collected from 51 subjects (41 cases and 10 Controls). Eligible participants were selected based on the availability of paired blood and surgical tissue specimens from cases suspected of breast cancer. We assessed the diagnostic and prognostic potential of a multi-gene expression panel developed by Geneverify Inc. using blood samples. The log₂-fold change in gene expression was calculated and used to generate a risk score for each subject. To determine the test's diagnostic performance and discriminatory power, we constructed receiver operating characteristic (ROC) and calibration curves. The Geneverify scores were compared against biopsy-confirmed results to calculate sensitivity and specificity. Geneverify scores were evaluated with respect to the patient's stage, grade, and metastasis. Results: A total of 51 subjects were evaluated, including 41 suspected breast cancer cases and 10 healthy controls. The Geneverify Dx genetic risk score effectively distinguished breast cancer patients from benign controls. The area under the ROC curve (AUC), indicating acceptable discriminatory performance. Using a risk score cutoff of 17 yielded 100% sensitivity. Calibration analysis demonstrated that the probability of breast cancer increased progressively with higher Geneverify Dx scores, reaching 100% positivity at scores 35. Spearman's rank correlation revealed a significant positive association between the risk score and clinical stage, as well as pathological grade. Furthermore, metastatic cases exhibited significantly higher scores compared to non-metastatic cases. Benign subjects showed markedly reduced gene expression levels and substantially lower risk scores. Conclusions: This prospective clinical study represents the first real-time validation of a blood-based, cell-free mRNA Geneverify test for screening and early detection of breast cancer. The test demonstrated exceptional diagnostic precision, reduce reliance on invasive biopsies, and ultimately improve patient management.