Rt-lamp Sensitivity Increase Using Unpurified Biosamples

Simple SummaryContent extracted from patent full text and abstract with AI.

This patent describes a method for detecting specific nucleic acid sequences, such as those from pathogens, directly from unpurified biological samples using an enhanced RT-LAMP (Reverse Transcription Loop-Mediated Isothermal Amplification) process. The key innovation is the use of specially modified primers (incorporating locked nucleic acids, LNAs) and optimized lysis buffer conditions, which allow accurate, sensitive nucleic acid amplification without the need for traditional RNA purification or expensive enzymes. This simplification enables fast, cost-effective, and scalable testing of diseases like COVID-19 and influenza directly from swabs or body fluids.

Use CasesContent extracted from patent full text and abstract with AI.

• Rapid, on-site diagnosis of infectious diseases (e.g., COVID-19, influenza, RSV) from patient swabs, saliva, or other body fluids.
• Population-scale disease screening in community settings, airports, workplaces, or schools.
• Molecular surveillance for pandemic preparedness and containment.
• Multiplexed detection of multiple pathogens or pathogen variants in a single sample/reaction.
• Field or resource-limited diagnostic testing in remote or developing regions.
• Automated, high-throughput testing in clinical laboratories.
• Fast identification of specific pathogen variants (e.g., SARS-CoV-2 variants of concern) via targeted sequencing post-amplification.

BenefitsContent extracted from patent full text and abstract with AI.

• Eliminates the need for laborious and costly RNA extraction steps, reducing complexity and turnaround time.
• High sensitivity and specificity for pathogen detection even with unpurified or minimally processed samples.
• Significantly reduces reagent and enzyme costs by omitting the need for separate thermostable reverse transcriptase enzymes.
• Compatible with automation and high-throughput workflows (e.g., robotics and 96-well plates), fostering scalable testing.
• Multiplexing capability allows detection of several pathogens or their variants in one reaction, streamlining differential diagnosis.
• Use of common and inexpensive buffer ingredients makes the method more widely accessible, particularly in resource-constrained settings.
• Minimizes contamination risk and cross-sample interference with novel sample handling hardware (e.g., piercing funnel device).
• Adaptable for point-of-care, decentralized, or portable testing scenarios.

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Inventors & Applicants

Patent Abstract

The present invention relates to a method for the detection of a target nucleic acid sequence in a sample, wherein the method comprises: a) optionally contacting the sample with a lysis buffer under conditions wherein the sample is lysed, b) optionally heating the lysis buffer; c) subjecting the lysate as obtained after step a) and optionally after step b) or the sample to an isothermal amplification reaction at a temperature of 30 to 75°C, preferably 45 to 75°, more preferably 60 to 70°C, even more preferred about 65°C with at least two primers specifically amplifying the target nucleic acid sequence, wherein the nucleotides of at least one primer comprise at least two locked nucleic acids (LNAs) which are not directly adjacent to each other within the nucleotides of the at least one primer, and wherein the isothermal amplification reaction comprises a DNA polymerase with reverse transcriptase activity and strand displacement activity and does not comprise a thermostable reverse transcriptase; and d) detecting the presence of the target nucleic acid sequence in the amplification product obtained after or during step d).