Nucleic Acid Extraction from Malleable Oral Polymers
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Solution Overview
Problem
There is a need for effective methods to detect and quantify microorganisms, such as bacteria, viruses, and fungi, that adhere to or are entrapped within polymers in the oral cavity, particularly in chewing gum, as existing methods are unclear about their ability to remove disease-causing organisms from the oral cavity.
Innovation Solution
A method involving the use of quantitative polymerase chain reaction (PCR) to extract and amplify nucleic acids from polymers that are malleable in the oral cavity, using organic solvents and buffered solutions to dissolve and separate nucleic acids from polymers like chewing gum, allowing for the identification and quantification of microorganisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to detect microorganisms in chewing gum, then the detection process is simple, but the ability to accurately detect and quantify disease-causing organisms is insufficient
Solution Approach 1:
The detection method is segmented into distinct stages: (1) extraction of nucleic acids from chewing gum using organic solvents and buffered solutions, (2) purification of extracted nucleic acids, (3) amplification using quantitative PCR, and (4) detection of amplified products. This segmentation allows each stage to be optimized independently, achieving high detection accuracy while maintaining procedural clarity.
Solution Approach 2:
Organic solvents and buffered solutions serve as intermediary substances that facilitate the extraction of nucleic acids from the polymer matrix of chewing gum. These intermediaries enable the separation of biological material from the gum base, making subsequent detection possible without directly modifying the detection apparatus.
2Loss of information
If nucleic acids are extracted from polymers using organic solvents and buffered solutions, then microorganisms can be detected and quantified, but the extraction process becomes more complex
Solution Approach 1:
The extraction process utilizes parameter changes in the polymer matrix by adjusting pH and solvent composition to optimize nucleic acid release from the gum base. buffered solutions with specific pH ranges facilitate the dissolution of polymer-bound nucleic acids, while organic solvents enhance extraction efficiency through selective solubility properties.
Solution Approach 2:
The extraction system employs a composite approach combining organic solvents (e.g., phenol, chloroform) with aqueous buffered solutions to create an optimized extraction environment. This composite solvent system addresses the dual challenge of breaking down the polymer matrix while preserving nucleic acid integrity, achieving high recovery without requiring multiple separate extraction steps.
3Measurement precision
If quantitative PCR is used to amplify and detect nucleic acids, then microorganism quantification is accurate, but the time and resources required increase
Solution Approach 1:
Nucleic acid extraction and purification are performed as preliminary actions before amplification. By completing the extraction and purification steps beforehand, the subsequent PCR amplification can proceed efficiently without interruptions, reducing total analysis time while maintaining quantification accuracy.
Solution Approach 2:
The methodology maintains continuous useful action through optimized workflow design where extraction, purification, and amplification steps are sequentially connected without unnecessary interruptions. The use of automated extraction systems and streamlined protocols ensures that each step transitions smoothly to the next, minimizing idle time while preserving the precision of quantitative detection.
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
This method enables the efficient extraction and quantification of microorganisms from polymers, providing insights into oral health and the potential for diagnosing infections or determining susceptibility to oral diseases, with the ability to detect a wide range of pathogens including Streptococcus mutans and Candida species.
Implementation Method 1
contacting the polymer with i) an organic solvent and ii) buffer solution
Implementation Method 2
separating the organic solvent and a buffer solution, wherein nucleic acid extracted from the polymer are contained within the buffer solution
Implementation Method 3
using an amplification method such as quantitative polymerase chain reaction (PCR)
Data Source
Figure 1
AI summary
The invention is directed to methods of extracting nucleic acids from microorganisms or mammalian cells adhered to polymers that are malleable within a living organism and particularly malleable in the oral cavity of the living organism. The invention also provides for method of detecting and quantitating microorganisms that adhere to malleable polymers, such as chewing gum.