Isothermal Amplification Primer Design for Rapid Point-of-Care Diagnostics

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Solution Overview

Problem

Current molecular diagnostics for antibiotic prescriptions are slow, costly, and not suitable for point-of-care settings, leading to over-prescription and misuse of antibiotics, which contributes to antimicrobial resistance and unnecessary side effects.

Innovation Solution

The use of Reverse-Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) with specific primer combinations for rapid and accurate amplification of biomarker mRNAs in clinical settings, allowing for quick discrimination between bacterial and viral infections.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Speed

If qRT-PCR is used for rapid biomarker measurement, then measurement speed is improved, but turnaround time remains >45 minutes which is still too slow for point-of-care

Engineering Contradiction:

Improvemeasurement speedVSAvoidturnaround time

Core Design Contradiction:

SpeedVSLoss of time

Solution Approach 1:

The patent replaces the thermal cycling mechanism of qRT-PCR with isothermal amplification (LAMP), eliminating the need for temperature cycling equipment and enabling faster, simpler amplification that can be completed in under 20 minutes without compromising measurement speed or accuracy

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the temperature parameter from cyclic variation (PCR) to constant isothermal condition (LAMP), which simplifies the amplification process, reduces equipment requirements, and accelerates the overall turnaround time while maintaining diagnostic accuracy

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If advanced molecular diagnostic technologies are used, then diagnostic accuracy is improved, but cost increases making them unsuitable for point-of-care settings

Engineering Contradiction:

Improvediagnostic accuracyVSAvoidcost

Core Design Contradiction:

Measurement precisionVSEase of manufacture

Solution Approach 1:

The patent employs disposable, pre-prepared LAMP reaction mixes and primers that are inexpensive to manufacture and use, eliminating the need for expensive instrumentation while maintaining high diagnostic accuracy through optimized primer designs and simple fluorescent detection

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

3Loss of time

If isothermal amplification is used for rapid diagnosis, then turnaround time is reduced, but amplification efficiency and accuracy may be compromised

Engineering Contradiction:

Improveturnaround timeVSAvoidamplification efficiency

Core Design Contradiction:

Loss of timeVSReliability

Solution Approach 1:

The patent performs preliminary optimization of LAMP primer sequences and reaction conditions during the development phase, pre-validating the amplification system to ensure high efficiency and accuracy are achieved under isothermal conditions before clinical deployment, thereby eliminating the need for time-consuming optimization during actual diagnosis

Inventive Principle:

Principle #10Preliminary action

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Enables rapid, accurate, and cost-effective diagnosis within 20 minutes, providing actionable insights for antimicrobial treatment decisions while maintaining high specificity and sensitivity, thus reducing antimicrobial misuse and improving patient outcomes.

Implementation Method 1

amplification of the subsequence is performed by Reverse-Transcription Loop-Mediated Amplification (RT-LAMP)

Methodology Applied

Scientific EffectReverse transcription:

Implementation Method 2

Reverse-Transcription Loop-Mediated Amplification (RT-LAMP) for the rapid, accurate, and efficient amplification of polynucleotides

Methodology Applied

Scientific EffectLoop-mediated isothermal amplification:

Data Source

PatentUS20240344130A1Isothermal amplification-based diagnosis and treatment of acute infection

Publication Date: 2024.10.17 INFLAMMATIX INC

AI summary

This invention provides primers and primer combinations that function efficiently in LAMP-based and/or other amplification systems and are useful for the diagnosis and subsequent treatment of acute infectious disease.