Steroid Surfactant and Cyclodextrin Nucleic Acid Extraction

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current nucleic acid extraction and amplification methods face challenges in efficiently extracting nucleic acids from microorganisms and viruses, particularly those with envelopes or single-stranded RNA, due to inhibitory effects from surfactants, which hinder rapid and sensitive detection in clinical infectious disease diagnostics.

Innovation Solution

A reagent system utilizing a surfactant with a steroid skeleton, such as sodium cholate, in combination with γ-cyclodextrin derivatives, specifically 2-hydroxypropyl-γ-cyclodextrin, to minimize inhibitory effects and facilitate efficient nucleic acid extraction and amplification, allowing for rapid and sensitive detection of pathogens.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a surfactant is used to extract nucleic acid from microorganisms, then extraction efficiency is improved, but the surfactant inhibits nucleic acid amplification reaction

Engineering Contradiction:

Improvenucleic acid extraction efficiencyVSAvoidinhibitory effect on amplification reaction

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

γ-cyclodextrin acts as an intermediary substance that selectively binds to the surfactant (bile acid or its derivatives) through host-guest inclusion complex formation. This mediation allows the surfactant to perform its extraction function while being sequestered by cyclodextrin, preventing it from inhibiting the subsequent PCR amplification reaction. The cyclodextrin effectively transfers the surfactant from an harmful state to a controlled complex state.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The harmful surfactant component is extracted or removed from the reaction system by forming an inclusion complex with γ-cyclodextrin. The cyclodextrin selectively takes out the surfactant molecules from the solution, concentrating them in a separate complex form that does not interfere with DNA polymerase activity, thereby eliminating the inhibitory effect while preserving extraction benefits.

Inventive Principle:

Principle #2Taking out (Extraction)

2Reliability

If conventional extraction methods are used, then nucleic acid can be obtained, but the process is time-consuming and lacks sensitivity for rapid detection

Engineering Contradiction:

Improvedetection sensitivityVSAvoidextraction and amplification time

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The extraction and amplification steps are merged into a single integrated reaction system. The surfactant-cyclodextrin system allows both extraction and PCR amplification to occur in the same tube without intermediate purification steps. This merging eliminates time-consuming transfer and cleanup operations while maintaining high detection sensitivity through the synergistic action of surfactant-enhanced extraction and cyclodextrin-protected amplification.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The surfactant performs preliminary extraction action on nucleic acids from microorganisms before the amplification reaction begins. By pre-extracting and releasing nucleic acids into the solution using surfactant action, the system prepares the sample in advance for immediate amplification, eliminating the need for separate extraction and purification steps that would consume time.

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

The proposed method significantly reduces the inhibitory effects of surfactants on nucleic acid amplification, enabling convenient and efficient extraction and amplification of nucleic acids from microorganisms and viruses, particularly those with envelopes or single-stranded RNA, thereby enhancing the sensitivity and speed of pathogen detection.

Implementation Method 1

γ-cyclodextrin having a C1-4 hydroxyalkyl group is contacted with a surfactant, thereby minimizing an inhibitory effect of the surfactant on a nucleic acid amplification reaction

Methodology Applied

Scientific EffectInclusion complex formation: Absorption (physical)

Implementation Method 2

A method is provided for extracting a nucleic acid of a nucleic acid extraction target from a sample containing the nucleic acid extraction target using a surfactant with a steroid skeleton

Methodology Applied

Scientific EffectSurfactant action: Surfactant

Data Source

PatentEP3597745B1Reagent for extracting and amplifying nucleic acid

Publication Date: 2022.08.24 TOSOH CORP

AI summary

The present invention addresses the problem of providing a reagent for extracting/amplifying a nucleic acid of a nucleic acid extraction target, the reagent being characterized in that a nucleic acid is conveniently extracted quickly and efficiently from the nucleic acid extraction target and inhibition of a nucleic acid amplification reaction is minimized, and the problem of providing a method for extracting or amplifying a nucleic acid using said reagent. The problems are solved by using a kit for extracting and amplifying a nucleic acid of a nucleic acid extraction target from a sample containing the nucleic acid extraction target, the kit including (i) a nucleic acid extraction reagent containing at least a surfactant having a steroid skeleton, (ii) γ-cyclodextrin having a C1-4 hydroxyalkyl group, and (iii) a nucleic acid amplification reagent.