100+
Hyperplex
Detect more than 100 targets in each sample instantly without sequencing (true multiplexing).
Powered by our novel Nanopixels™.
SNR
Single-nucleotide resolution
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.
Ultra-sensitive
1 copy
Reach extreme sensitivty while retaining single molecule specificity, surpassing qPCR using our novel hpPCR assay.
Fast & Simple
4h
Results within 4h instead of weeks for NGS. Easy workflow, with few steps using our kits & consumables.
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Hands on time ~30 min from extracted sample.
Sensitivity, specificity, multiplexity. No compromises.
STANDARD EQUIPMENT
No need for advanced equipment. Use standard lab equipment, read out the results in a fluorescent microscope within 3h.
COST-EFFECTIVE
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.
MOLECULAR COUNTING
Hyperplex PCR is a digital molecular assay where targets molecules are amplified and counted individually, enabling unprecedented quantiative accuracy and VAF sensitivity.
TECHNOLOGY
WHAT IS hpPCR™?
Hyperplex 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 microscopes.
HOW IT WORKS
Sample
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.
PCR Amplification
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.
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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 Nanopixels™, capable of generating 100+ synthetic colors using common fluorescence filter sets.
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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.
Image Analysis
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 molecules in the sample - giving an intrinsically digital read-out without the need for compartmentalization as in digital PCR.
Throughput
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Each microscope slide can fit multiple samples (up to 16), and each sample now contains a full picture of your experiment or analysis.
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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.