Rapid Nucleic Acid Extraction Using Reducing Agents for Diagnostics
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Solution Overview
Problem
Current methods for diagnosing pathogenic infections like Chlamydia trachomatis or Neisseria gonorrhoeae require time-consuming nucleic acid purification procedures, trained users, and expensive equipment, making them complicated and less accessible for rapid diagnostic tests.
Innovation Solution
A method involving a reducing agent like dithiothreitol (DTT) and a buffer at ambient temperature for up to 30 minutes to extract nucleic acids directly from samples, followed by immediate nucleic acid amplification without additional purification steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If nucleic acid purification procedures are performed prior to amplification, then diagnostic accuracy is improved, but diagnostic time and procedural complexity increase
Solution Approach 1:
The patent combines the nucleic acid extraction and amplification steps into a single integrated reaction tube. The extraction reagent mixture containing reducing agents (DTT or β-ME) and buffer is prepared in advance, and the clinical sample is directly added to this mixture for incubation. After incubation at ambient temperature for up to 30 minutes, the same mixture is used directly for nucleic acid amplification without any separation or purification steps, thereby eliminating time-consuming intermediate procedures while maintaining diagnostic accuracy.
Solution Approach 2:
The extraction reagent mixture is prepared in advance before the diagnostic test is performed. The reducing agents and buffer are pre-combined in appropriate concentrations, creating a ready-to-use extraction system. This preliminary preparation allows the actual diagnostic procedure to proceed rapidly when the clinical sample is introduced, reducing the overall diagnostic time while ensuring proper extraction conditions are already in place.
2Reliability
If nucleic acid purification procedures are performed prior to amplification, then nucleic acid quality is improved, but device complexity and user training requirements increase
Solution Approach 1:
The patent merges multiple steps (extraction, purification, and amplification preparation) into a single integrated procedure performed in one reaction tube. The extraction reagent mixture with reducing agents creates conditions that both extract nucleic acids from the clinical sample and maintain their quality for subsequent amplification, eliminating the need for separate purification equipment and procedures.
Solution Approach 2:
The extraction reagent mixture self-performs the purification function through the chemical action of reducing agents that lysed bacterial cells and release nucleic acids while the buffer maintains appropriate pH and ionic conditions. This self-service mechanism eliminates the need for complex external purification equipment or highly trained operators, making the procedure accessible to clinicians and even patients with minimal training.
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 and efficient nucleic acid extraction and amplification, allowing for quicker diagnostic results with ease and reduced need for specialized equipment, suitable for both clinicians and patients.
Implementation Method 1
combining the sample with an extraction reagent, the reagent comprising a reducing agent
Data Source
AI summary
Disclosed herein are novel methods and compositions for rapidly extracting and amplifying nucleic acids from a sample where the sample is combined with an extraction reagent comprising a reducing agent to form a mixture and incubating said mixture at ambient temperature for a period of time not exceeding 30 minutes to generate a nucleic acid extract. In certain embodiments of the method, the nucleic acid extract is subjected to a nucleic acid amplification reaction. In certain aspects, oligonucleotide primers specific for nucleic acids of Chlamydia species and/or Neisseria species are added prior to initiating the amplification reaction.