rRNA Assay for Rapid Bacterial Density Estimation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for estimating microbial density in specimens, particularly clinical specimens, face challenges due to variability in rRNA copies per cell and unknown growth phases, making accurate determination of bacterial density difficult and time-consuming.
Innovation Solution
A method involving an rRNA assay to determine bacterial rRNA concentration, followed by conversion to a bacterial density value using a predetermined correlation, allowing for rapid and accurate estimation of microbial density in specimens, even when rRNA copies per cell vary and growth phase is unknown.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If rRNA quantification is performed to estimate bacterial density, then measurement speed is improved, but measurement precision deteriorates due to variability in rRNA copies per cell and unknown growth phases
Solution Approach 1:
The patent transforms the measurement parameter from direct bacterial density estimation to rRNA concentration measurement. By detecting rRNA concentration through hybridization signals and using a predetermined correlation function, the system achieves rapid measurement while accounting for rRNA variability through established correlation relationships between rRNA concentration and bacterial density across different growth phases
Solution Approach 2:
The patent introduces rRNA concentration as an intermediary parameter between direct bacterial detection and density estimation. The hybridization signal detects rRNA concentration, which then serves as a mediator to estimate bacterial density through a predetermined correlation, bridging the gap between rapid rRNA measurement and accurate bacterial density determination
2Measurement precision
If conventional bacterial density methods are used to account for rRNA variability, then measurement precision is improved, but time consumption increases
Solution Approach 1:
The patent performs preliminary work by establishing a predetermined correlation function between rRNA concentration and bacterial density before actual measurements. This pre-established correlation accounts for rRNA variability across different growth phases, allowing rapid direct estimation without time-consuming culturing or growth phase determination during the actual measurement process
Solution Approach 2:
The patent uses rRNA molecules as copies or proxies for bacterial cells. Instead of directly counting or culturing bacteria, the system detects rRNA concentration which serves as a molecular copy representing bacterial presence and density, enabling rapid measurement without traditional time-consuming bacterial enumeration 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 reliable and rapid estimation of microbial density in clinical specimens, facilitating timely clinical decisions and optimizing antimicrobial susceptibility testing, with results available in under an hour or less.
Implementation Method 1
contacting the specimen with an oligonucleotide probe that specifically hybridizes with a target nucleic acid sequence region of ribosomal RNA
Implementation Method 2
a magnetic lysis blade and lysis beads are disposed in each of the lysis chambers and a plurality of stationary magnets are disposed adjacent to and separate from the microfluidic CD. The stationary magnets are configured to magnetically interact with each of the magnetic lysis blades upon rotation of the microfluidic CD
Data Source
AI summary
A method of determining a bacterial density in a specimen may include the steps of: (a) conducting an RNA assay on the specimen to determine a microbial rRNA concentration, wherein the microbial rRNA concentration is defined as the number of rRNA molecules per volume of the specimen; and (b) converting the rRNA concentration to a bacterial density value.