molecular dx

The 2PG MoMâ„¢ can detect and quantify DNA/RNA targets across a range of concentrations.

Silicon nanopores: single molecule sensors

Silicon nanopore sensors electrically sense individual molecules, one at a time. No optics. No chemistry.

Voltage pulls molecules through, impeding current by a measured amount.

Matrix contains DNA, RNA, proteins, small molecules, resulting in many different event signatures. Single molecule “event” has depth and duration, proportional to its size. Events are plotted based on impedance (depth) and duration.

Detecting amplified genomic markers

Goal: Screen saliva sample spiked with HCV at clinical concentration levels (~500 copies per ml)

  • Swabbed sample is placed into test strip is inserted into amplifier for in situ HCV amplification (7 mins).
  • Amplified product is detected alongside a synthetic calibrant and data is analyzed in real time

HCV positive sample (blue) and HCV negative sample (black) show clear detection of the positive sample above background.

Multiplexed SNP detection

Method: Estimating the fractional abundance of KRAS mutation G12D transcripts compared to KRAS wild-type transcripts.

  1. Post cfDNA extraction, target sequence is amplified using minimal PCR cycles.
  2. SNP specific probes are added and allowed to bind.
  1. Tagged amplicons (WT and mutant) are simultaneously detected in the pore. The different sizes are classified by their distinct event populations for mutant (black) and wild-type (blue), with false positives (<0.5%).

Different probes produce different impedance signatures

Assay demonstrates:

  • Multiplexing
  • SNP detection using probe-payloads
  • Fast assay time
  • PCR unbiased amplification method achieves rare target detection

Multiplexing multiple targets: Chlamydia, Gonorrhea, Trichomonas

Raw sample contains Chlamydia, Gonorrhea, Trichomonas (100 copies), among other background.

Amplify all DNA together in the same well. Each primer set contains different sized probes.

Post-amplification, 100bp amplicons are tagged with different sized probes.

Probes cause amplicons to produce unique signal populations. Scatter plot shows each target is present in the sample, demonstrating three-plex.