Detect more than 100 targets in each sample instantly without sequencing (true multiplexing).
Powered by our novel hyperprobes.
Detect RNA/DNA sequences with single nucleotide specificity. Primer design is straight-forward with our novel assay and new panels can be generated swiftly for custom targets.
Reach extreme sensitivty while retaining single molecule specificity, surpassing qPCR using our novel hpPCR assay.
Fast & Simple
Results within 3h instead of weeks for NGS. Easy workflow, with few steps using our kits & consumables.
Hands on time ~30 min from extracted sample.
Sensitivity, specificity, multiplexity. No compromises.
No need for advanced equipment. Use standard lab equipment, read out the results in a fluorescent microscope within 3h.
Our revolutionary technology removes the need for complex instrumentation & consumables such as patterned flow-cells or microfluidics, enabling a completely new scale of hyperplexing for every lab, and for massive scale.
Detect 1 : 100 000 mutation.
Our assay enables ultra-sensitivity while retaining specificity allowing for sensitive VAF analysis. Combined with the ability to target more than 100 markers, we expect to revolutionize cancer diagnostics.
WHAT IS hpPCR?
Hyperplexed Padlock PCR (hpPCR) is a massively multiplexed, highly specific and ultra-sensitive molecular detection platform packaged into a simple and efficient workflow. The hpPCR platform can be used in any lab equipped with regular PCR cyclers and a fluorescence microscope.
HOW IT WORKS
DNA or RNA is extracted from the sample, such as wastewater, crops, food, blood, saliva, bone marrow or tissue. A broad set of target sequences (100+) can be analyzed in the same sample without splitting or sequencing.
The DNA sequences of interest, e.g. mutations in a patient’s malignant cells, a panel of DNA sequences for diagnosing disease, or a panel of pathogens, are first enriched by a multiplexed PCR amplification.
Specific Probing & RCA Amplification
The enriched targets are then probed with multiple target specific barcoded padlock-probes (PLPs) which undergoe a ligase-mediated circularization.
Circularization is only successful upon perfect sequence match, giving the ability to discriminate mutations with single-nucleotide specificity and eliminate false-positives from PCR artifacts.
The circularized strands containing the target region together with a unique barcode are then amplified by Rolling Circle Amplfication (RCA) to generate micrometer sized RCA products (RCPs).
Hyperplex Labelling & Imaging
The amplified targets are captured on a surface (e.g. microscope slide) and labelled with APLEX Bio's proprietary fluorescent hyperprobes, capable of generating 100+ synthetic colors using common fluorescence filter sets.
The detection of all targets is performed simultaneously using a fluorescence microscope with low magnification (10-20x) and 4-5 channels in a single step.
No sample splitting or cycling needed, from start to end.
RCPs are then identified as bright fluorescent dots, each a single DNA target molecule. APLEX Bio's proprietary software identifies each unique target based on its corresponding color code, and counts the amount of dots in the sample - giving an intrinsically digital read-out without the need for compartmentalization as in digital PCR.
Each microscope slide can fit multiple samples (up to 16), and each sample now contains a full picture of your experiment or analysis.
For even higher throughput, 96-well or 384-well plates can be used, and thanks to the hyperplexing in each sample a massive throughput can be achieved in comparison to other PCR technologies.