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ProbeLAMP^TM

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ProbeLAMP^TM

Displaceable Probe LAMP: ProbeLAMP^TM

Results in minutes. Inexpensive equipment. Test in public spaces.

Problem

As COVID-19 proved, pathogens spread rapidly because the public lacks a test that can be used at entrances to public spaces to identify virus carriers. Existing isothermal assays are too slow, too insensitive, and difficult to multiplex. An effective test must be rapid, safe, and cheap, with several targets tested at the same time.

Standard LAMP needs no heat cycling and produces results rapidly. But it has only inconvenient readout. ProbeLAMP solves these problems.

Solution

Create an amplification-detection architecture with fluorescent glow arising from separation of a fluorescent group from a fluorescence quencher.
Run test at 65 °C killing the target virus.

Impact

Faster, cheaper detection of targets of interest:

Tests that detect both the pathogen target and a control
Easy visualization; orthogonal fluorescence
Low cost instrument being developed by Sparsek

Chemistry behind ProbeLAMP^TM

We invented a test that:

Uses LAMP architecture to get amplification going
Runs at 65°C, which inactivates the target virus
Signals, by a displaceable probe, a short fluorescently tagged DNA that binds to a DNA molecule with a quencher, which makes the fluor dark
Fluorescent DNA is displaced from quenching DNA during amplification
That complementary oligonucleotide has a 5'-quencher, thus each probe is delivered to the amplification mixture as a target-sequence-independent double-strand probe region and a single-stranded target-priming region

When target pathogen is absent, no fluorescence is seen. The fluorophore is quenched by the nearby quencher. But as amplification proceeds, in the LAMP process, a primer extends into the fluor:quencher duplex, displacing the fluor from the rest of the amplification product. Free to move away from the quencher, the fluor now glows, signalling the presence of the virus in real-time.

Publications

Loop creation

LAMP architecture allows creation of a looped "dumbbell" structure for isothermal amplification.

Amplification

The dumbbell structure is used as a template for repeated amplification.

Fluorescence

When amplified target is present, the quencher is displaced by polymerase causing fluorescence.

Probe displacement

Fluorescently tagged probe is displaced by polymerase, allowing for a visible test result.