Multiplexed Fuel Analysis for Rapid Microbial DNA Detection
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Solution Overview
Problem
Current methods for detecting microbial contamination in fuels are time-consuming, complex, and limited in their ability to identify a broad spectrum of microbes, requiring significant manual manipulation and expertise, and are not suitable for rapid, field-deployable assays that can provide actionable results within two hours.
Innovation Solution
The development of multiplexed Rapid DNA assays that allow for the simultaneous detection of multiple microbial species and strains in fuel samples using microfluidic chips and rapid thermal cyclers, enabling rapid DNA purification and PCR amplification, with the ability to identify specific organisms and strains in various fuel types.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional microbial detection methods are used, then detection accuracy can be maintained, but the analysis time is excessively long and manual manipulation is required
Solution Approach 1:
The patent combines multiple detection functions into a single microfluidic chip that performs DNA extraction, PCR amplification, and multiplexed detection simultaneously. This integration eliminates the need for separate manual操作步骤 and reduces overall detection time while maintaining accuracy through automated process control.
Solution Approach 2:
The patent replaces manual mechanical operations with automated microfluidic systems. The microfluidic chip uses integrated pumps, valves, and reaction chambers to automate DNA extraction and amplification processes, eliminating time-consuming manual manipulation while increasing throughput through parallel processing.
2Adaptability or versatility
If broad-spectrum microbial detection is implemented, then detection capability is improved, but assay complexity increases significantly
Solution Approach 1:
The patent employs a universal microfluidic chip design that can detect multiple microbial species and strains simultaneously through multiplexed PCR assays. The same chip platform can be configured with different primer sets to detect various targets, providing broad detection capability without requiring multiple separate assays or complex equipment.
Solution Approach 2:
The patent segments the detection process into distinct functional modules within the microfluidic chip, including sample preparation, DNA extraction, PCR amplification, and detection zones. Each module performs a specific function, allowing complex multiplexed assays to be broken down into manageable steps that can be automated and controlled independently.
3Speed
If rapid DNA assays are used, then detection speed is improved, but the ability to identify specific strains may be compromised
Solution Approach 1:
The patent uses periodic thermal cycling in the PCR process to selectively amplify specific DNA sequences. The rapid thermal cycler performs repeated heating and cooling cycles that denature DNA, allow primer annealing, and extend new strands, enabling specific strain identification through sequence-specific amplification within a rapid time frame.
Solution Approach 2:
The patent uses PCR amplification to create multiple copies of specific DNA sequences from the microbial samples. This copying process amplifies trace amounts of target DNA to detectable levels, enabling accurate strain identification through sequence analysis while maintaining rapid detection through efficient in vitro synthesis rather than time-consuming culture methods.
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, efficient, and accurate identification of microbial contaminants in fuels, allowing for precise localization of contamination sources, forensic analysis of fuel origin, assessment of remediation efficacy, and detection of indicator species for exploration, all within a few hours.
Implementation Method 1
rapid DNA purification
Implementation Method 2
DNA purification
Implementation Method 3
rapid thermal cyclers, enabling rapid DNA purification and PCR amplification
Implementation Method 4
PCR amplification
Implementation Method 5
multiplexed Rapid DNA assays that allow for the simultaneous detection of multiple microbial species
Data Source
AI summary
Compositions, testing chambers and methods for testing a fuel sample for microbial contamination (including fuels treated with a biocide) are provided, which comprise: a quantity of hydrocarbon fuel; a microbial contamination wherein the microbial contamination further comprises nucleic acid in the form of both DNA, RNA or a combination thereof, and an analyzing solution; wherein the analyzing solution comprises at least six (6) primer pairs for amplification of at least one target locus, wherein at least one primer of each pair of primers is labeled with a fluorescent dye and wherein at least one of the primer pair binds to the nucleic acid of the microbial contamination.