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INTRODUCING
HYPERPLEX PCR 
hpPCR

100+

Hyperplex

Detect more than 100 targets in each sample instantly without sequencing (true multiplexing).
Powered by our novel Nanop
ixels™.

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.

Hands on time ~30 min from extracted sample.

Specs

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.

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hpPCR

HOW IT WORKS

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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.

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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.

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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 Nanopixels™, 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.

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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.

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Throughput

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.

Ready to upgrade your PCR?

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