Microbial Population Analysis via Primer-Based Fingerprinting

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Solution Overview

Problem

Current methods for analyzing populations of microorganisms, such as bacterial populations, are laborious, time-consuming, and limited in their ability to provide reliable and reproducible data on the overall composition of complex microbial communities, especially when dealing with unknown or diverse bacterial populations.

Innovation Solution

A method involving specific primer sets that target conserved regions across different taxonomic groups, allowing for amplification and detection of fragments with differences in size, number, and nucleotide sequence, enabling a 'fingerprint' analysis of microbial populations without requiring detailed identification of individual organisms.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If traditional methods are used to analyze microbial populations, then detailed identification of individual organisms can be achieved, but the analysis becomes extremely laborious and time-consuming

Engineering Contradiction:
Improveidentification accuracyVSAvoidanalysis time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent segments the complex task of identifying individual microorganisms into a simplified approach that analyzes population-level patterns. Instead of identifying each organism separately (which is time-consuming), the method divides the problem into: (1) extracting DNA from the entire population, (2) amplifying specific genetic regions with primers, and (3) analyzing the collective fragment patterns. This segmentation allows rapid population analysis without the laborious step of individual organism identification.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent extracts only the essential information needed for population analysis - specific DNA fragments from conserved genetic regions - rather than attempting to identify and characterize every individual organism. By taking out and amplifying only the relevant genetic markers using targeted primers, the method obtains sufficient data for population characterization while avoiding the time-consuming process of complete organism identification.

Inventive Principle:
Principle #2Taking out (Extraction)

2Loss of information

If comprehensive analysis of complex microbial communities is performed, then detailed composition data can be obtained, but the method becomes too laborious for routine application

Engineering Contradiction:
Improvepopulation composition dataVSAvoidmethod simplicity
Core Design Contradiction:
Loss of informationVSEase of manufacture

Solution Approach 1:

The patent creates a universal method that can analyze diverse microbial populations using the same standardized procedure. The primer sets are designed to target conserved genetic regions present across different taxonomic groups, allowing the same methodology to be applied universally to bacteria, archaea, and other microorganisms. This multi-functionality enables routine application across different samples and environments without requiring complex, sample-specific protocols.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent changes the analytical parameter from individual organism identification to population-level fragment pattern analysis. By shifting from identifying specific organisms to analyzing the collective characteristics of DNA fragments (size, abundance, composition), the method maintains comprehensive population data while dramatically simplifying the procedure for routine application.

Inventive Principle:
Principle #35Parameter changes

3Reliability

If detailed identification of individual microorganisms is attempted, then complete population characterization is achieved, but reproducibility and reliability are reduced due to complexity

Engineering Contradiction:
Improvedata reproducibilityVSAvoidmethod complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent performs preliminary action by targeting and amplifying specific conserved genetic regions before analysis. The primer sets are designed in advance to bind to highly conserved sequences across taxonomic groups, ensuring that the same genetic markers are analyzed in every sample. This preliminary targeting of standardized regions before population analysis enhances reproducibility by eliminating variability introduced by attempting to identify diverse individual organisms with different genetic characteristics.

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

This method allows for a reliable, non-laborious analysis of microbial populations, providing information on taxonomic groups and relative amounts, and can detect alterations over time, enhancing sensitivity and reproducibility, and is applicable to various environments like the gastrointestinal tract and water systems.

Implementation Method 1

performing an amplification reaction using said primer sets, thereby producing fragments having detectable differences in size, number, nucleotide sequence and/or label

Methodology Applied
Scientific EffectPCR amplification:

Data Source

PatentEP2152904B1Microbial population analysis
Publication Date: 2017.11.22 MICROBIOME
  • EP2152904B1 patent drawingFigure 1
  • EP2152904B1 patent drawingFigure 2
  • EP2152904B1 patent drawingFigure 3

AI summary

The current invention relates to a method for analysis of a population of micro-organisms (e.g. bacterial population) of different taxonomic groups in an environment suspected to contain said bacteria, primers, primer sets and pair of primer sets suitable for use is such method, and use of such method in determining the effect of external factors like drugs, nutrients and pesticides on bacterial populations of different taxonomic groups.