79. Extraction of multiple analytes in liquid biopsy may improve the diagnosis of breast cancer

Aly Abdelkareem

Nafiseh Jafari

Nafiseh Jafari, Ph.D., is Chief Science Officer at nRichDX. Nafiseh joined nRichDX in January 2020 and has over ten years of experience in life science research. She is a scientific leader passionate about building and managing high-performance teams. Nafiseh has managed multimillion-dollar development projects leading to the successful creation and launch of multiple products. Her expertise in talent management and quality/design control systems makes her a vital member of the nRichDX team. Immediately before joining nRichDX, Nafiseh was a Project Manager leading cross-functional teams at Abbott Diagnostics. She holds a B.S. degree in Chemistry from Sharif University of Technology and earned her Ph.D. degree in Biochemistry and Molecular Biology at Western Michigan University.


Nafiseh Jafari, Mayer Saidian, Jason Saenz, Cory Arnold, Andrew Dunnigan, Carlos Hernandez, Lauren Lee

nRichDX, Irvine, CA, United States

Introduction: Since a single biomarker is inefficient in accurately identifying most cancers, integrated liquid biopsy using multiple biomarkers might be a promising method to facilitate early detection and treatment of cancer. This study demonstrates an optimized extraction method for cfTNA (Cell-Free Total Nucleic Acid) and circulating tumor cells (CTC) from a single blood sample.

Methods: Whole blood was collected from healthy donors and processed for CTC and cfTNA extractions. The cellular components were used to extract CTCs, while plasma was used to extract cfTNA. One hundred MCF7 cells were spiked into whole blood. Plasma was removed and spiked with 10ng/mL of DNA and RNA. The nRichDX Revolution System was used for all extractions. cfTNA percent recovery was calculated by a PIK3CA mutation detection RT-qPCR assay. Immunocytochemistry was used to assess the recovery of CTCs.

Results: Multiple analytes were successfully extracted from a single blood sample using the Revolution System. ICC staining showed positive recovery of EpCAM (+), CD45 (-), and DAPI (+), indicating CTC recovery. Extracted cfTNA contained the PIK3CA breast cancer mutation and was successfully amplified via RT-qPCR.

Conclusion: Recent studies show that multiple biomarker testing provides another level of biological information regarding the tumor and its microenvironment. Cancer profiling from a single blood draw can allow greater precision and efficiency for cancer diagnostics. A multi-technology, preanalytical approach that leverages high-efficiency epithelial CTC, cfDNA, and cfRNA extractions enables greater insight for clinicians, and drug development professionals focused on early cancer detection.