CIRCULOGENE’s molecular cancer diagnostics services leverage proven technology and provide a noninvasive and cost-effective solution for high-sensitivity monitoring across a variety of sample types. Our unique technology uses minimal volumes of blood to provide unprecedented tumor monitoring information with an up-to-date clinical status in a more complete, accurate, timely, less expensive and less invasive fashion.

  • CLIA-ready
  • Clinically validated assays
  • Unprecedented levels of cell-free DNA recovery from droplet volumes of blood translates into high sensitivity
  • Open platforms, Ion Torrent Proton for both mutation monitoring and profiling
  • Integrated workflow for consistent and reliable longitudinal studies

Clinical Challenges

Individualized cancer management for treating cancer patients with precision therapy is currently still challenged by the following limitations:

Tumor heterogeneity: tumors are heterogeneous and exhibit continuously polyclonal expansion, thus complicating diagnosis, treatment and the assessment of acquired resistance. Furthermore, tumor tissues are limited both spatially to the region biopsied and temporally to the state of tumor at the time of biopsy.

Accessibility of tumor tissue: the majority of patients treated with targeted therapies ultimately develop resistance, metastasis or recurrence. Effective monitoring with several tumor samples is not clinically practical with current, invasive biopsy techniques. Most importantly, tumor tissue may not be available or has been exhausted due to multiple testings.

Unmet Needs in Liquid Biopsy and CIRCULOGENE’s Solution

Novel cell-free DNA enrichment method for high-quantity and high-quality starting materials:
Circulating, cell-free DNA is highly fragmented and presented at very low concentrations. This makes its isolation challenging due to the requirements of high-volume input, costs and labor intensity. Our proprietary cell-free DNA enrichment process possesses the novel characteristic of low input (as low as 50 uL of plasma) and high output (>300 ng/mL), allowing ultra-high recovery and detection of tumor DNA directly from droplet volumes of unprocessed blood.

Advanced NGS technology for sensitive longitudinal mutation detection:
Detection of low-frequency mutations through periodic cell-free DNA analysis could predict tumor progression before the lesions are large enough to be detected by imaging. Analysis beyond a single mutation could also be warranted to capture tumor heterogeneity for effective treatment decision making. Liquid biopsies are not as spatially limited as tissue biopsies and can show a global spectrum of mutations that occur throughout tumor development in our body. The sensitivity of conventional analytical methods such as Sanger sequencing is not sufficient to detect low-frequency variants. Targeted deep sequencing by next-generation sequencing (NGS) provides a cost-effective alternative for high-throughput analysis of multiple mutations with high sensitivity.

Validated sample processing and analysis procedure in clinical practice:
The utility of cell-free DNA for clinical application demands the implementation of stringent pre-analytical validation of our laboratory developed tests to ensure consistent quality data acquisition in our CLIA-certified laboratory.

Key Metrics

  • Regions Analyzed: Mutation hotspot regions of 50 cancer-associated genes
  • Sequencing Method: Ion Proton NGS
  • Assay Sensitivity: >1.0%
  • Target Sequencing Coverage: 5,000x
  • Turnaround Time: 5-7 days
  • Sample Requirements: Finger stick, blood/microcontainer, purple top EDTA 250-500 uL tube
  • Sample Types: Plasma or Serum
  • Sample Input Required: Droplet Volumes (50-500 uL)

Sample Collection

Large sample volume, low yield and labor intensiveness are the major barriers to the routine use of cell-free DNA-based testing in current clinical settings. Specifically, cell-free DNA recovery efficiency using the current standard silica membrane/bead method is extremely low due to unavoidable loss in steps such as binding, washing and elution. This in turn results in a minimum sample requirement of 5-10mL of blood.

While almost all liquid biopsy providers are working and perfecting their novel technologies to selectively enrich or amplify tumor specific, cell-free DNA from a dominantly normal population, their starting materials are still those “incomplete” DNA molecules that survived after extraction/isolation. As a result, no matter how sensitive their technology is (downstream), they cannot detect cell-free DNA already lost during sample preparation (upstream). Further, there is uncertainty if even 10 mL of blood is enough.

In contrast, CIRCULOGENE focuses on the first step in the cell-free DNA process, making sure we capture all of the cell-free DNA molecules existing in the original sample. Our breakthrough, proprietary method enables recovery of both necrotic and apoptotic cell-death-associated, cell-free DNA, with a high-yield, high-degree genomic representation and high sequencing quality.

Sample Quality and Quantity

CIRCULOGENE’s proprietary method provides a simple, streamlined, and robust workflow optimized for cell-free DNA enrichment/recovery from a droplet of plasma or serum as an alternative to low-quantity, poor-quality DNA from challenging tissue biopsy samples. This sample preparation breakthrough enables multiple analyses on a droplet sample by a broad range of genomic platforms, including (NGS), microarrays, quantitative PCR (qPCR) and droplet digital PCR (ddPCR). Working with challenging and precious samples is no longer a barrier to personalized cancer therapy. Our technology allows us to work with a sample volume as small as 50 uL, saving the remainder of your biobank for other clinical testing.

We compared the sensitivity of our proprietary cell-free DNA enrichment method to the standard DNA extraction kit used industry wide today. Results demonstrated 100x more cell-free tumor DNA in the original sample using a single drop of blood.

Sensitivity and Accuracy

CIRCULOGENE’s cell-free DNA enrichment process is a complete solution for generations of high-yield, high-quality, well-represented tumor DNA from droplet amounts of plasma or serum. It is the first non-invasive method that allows accurate NGS genomic analyses directly from droplet volumes of blood. Our protocol can be applied to a broad range of clinical genetic tests with the advantages of minimal sample volume, maximal yield and streamlined workflow with reduced cost and turnaround time.

Automation

CIRCULOGENE’s automated solutions have been developed to establish a complete and efficient NGS workflow from sample-in to report-out. The simplicity and robustness of our automated process enables interfaces and cross-talks between a variety of platforms and laboratory information management systems. Using this simplified workflow, an automated workstation can easily process 96 or more samples at one time.