Multiplexed Fuel Microbe Assays Using Microfluidic PCR

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for detecting microbial contamination in fuels are time-consuming, labor-intensive, and limited in their ability to identify a broad spectrum of microorganisms, making them unsuitable for rapid, field-deployable assays that can detect specific microbial species and strains.

Innovation Solution

The development of multiplexed Rapid DNA assays that can simultaneously amplify and identify multiple microbial species and strains in fuel samples using microfluidic chips and rapid thermal cyclers, enabling rapid detection within two hours.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If traditional microbial detection methods are used, then detection accuracy for specific species is improved, but detection time and labor requirements increase significantly

Engineering Contradiction:

Improvedetection accuracyVSAvoiddetection time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The detection process is segmented into three integrated modules: DNA extraction module, multiplex PCR amplification module, and detection module. Each module handles specific tasks independently, enabling parallel processing of multiple microbial targets simultaneously, thus reducing overall detection time while maintaining species-specific accuracy

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention uses universal primers that can amplify DNA from multiple different microbial species (bacteria, fungi, algae) simultaneously in a single reaction. This multi-functional approach allows one assay to detect many different targets, reducing the number of separate tests needed while preserving detection precision for each species

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Adaptability or versatility

If traditional microbial detection methods are used, then comprehensive identification of microorganisms is improved, but operational complexity and labor intensity increase

Engineering Contradiction:

Improveidentification capabilityVSAvoidoperational simplicity

Core Design Contradiction:

Adaptability or versatilityVSEase of operation

Solution Approach 1:

Multiple detection functions are merged into a single multiplex PCR assay. The assay combines primers for different microbial targets (bacterial 16S rRNA, fungal 18S rRNA, algal ITS regions) in one reaction tube, allowing simultaneous identification of multiple organism types without requiring separate operational procedures for each

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The system uses self-priming oligonucleotides with built-in fluorescent labels that automatically bind to complementary DNA sequences during amplification. This self-service mechanism eliminates the need for manual post-PCR analysis or complex interpretation steps, reducing labor while maintaining comprehensive identification capability

Inventive Principle:

Principle #25Self-service

3Productivity

If rapid detection methods are developed, then detection speed is improved, but ability to detect broad spectrum of microorganisms deteriorates

Engineering Contradiction:

Improvedetection speedVSAvoidspectral coverage

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The invention employs periodic thermal cycling (denaturation, annealing, extension) in the PCR process, where each cycle rapidly amplifies specific DNA sequences. This periodic action allows the system to quickly cycle through multiple amplification rounds in parallel, maintaining high detection speed while the multiplex design ensures broad spectral coverage of different microbial types

Inventive Principle:

Principle #19Periodic 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

Enables rapid, field-deployable detection of a wide range of microbial species and strains in fuels, facilitating effective prevention and remediation strategies, tracing contamination sources, and assisting in fuel exploration.

Implementation Method 1

rapid thermal cyclers

Methodology Applied

Scientific EffectThermal cycling: Temperature Gradient

Implementation Method 2

simultaneously amplify and identify multiple microbial species

Methodology Applied

Scientific EffectPCR amplification:

Implementation Method 3

microfluidic chips

Methodology Applied

Scientific EffectCapillary electrophoresis: Capillary Electrophoresis

Data Source

PatentUS12480166B2Multiplexed fuel analysis

Publication Date: 2025.11.25 ANDE CORP

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.