Multiplex PCR Assay for Sepsis Pathogen Identification
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Solution Overview
Problem
Current methods fail to efficiently identify causative bacteria of sepsis from multiple pathogens, which is crucial for timely treatment of severe infections leading to sepsis.
Innovation Solution
A method using PCR with specific primers and probes targeting the gyrB gene of Escherichia coli, the nuc gene of Staphylococcus aureus, the atlE gene of Staphylococcus epidermidis, the gyrB gene of Klebsiella pneumoniae, and the rpoB gene of Enterococcus spp. for simultaneous identification of causative bacteria in sepsis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple separate identification methods are used for different bacteria, then each bacterium can be identified accurately, but the identification process becomes time-consuming and complex
Solution Approach 1:
The patent combines multiple separate bacterial identification methods into a single multiplex PCR system. Different primer sets targeting specific bacterial genes ( gyrB for E. coli and Klebsiella, nuc for S. aureus, atlE for S. epidermidis, rpoB for Enterococcus) are merged into one reaction mixture, allowing simultaneous amplification and detection of multiple pathogens in a single test, thereby reducing identification time while maintaining accuracy
Solution Approach 2:
The invention creates a universal identification system that can detect multiple types of bacteria causing sepsis using a single PCR platform. The method employs universal PCR conditions and detection protocols that work across different bacterial targets, making the system multi-functional for identifying various sepsis-causing pathogens without requiring separate specialized tests for each bacterium
2Measurement precision
If multiple separate identification methods are used for different bacteria, then each bacterium can be identified accurately, but the overall procedure becomes complex
Solution Approach 1:
The patent merges multiple identification procedures into a single multiplex PCR assay. By combining different primer sets and detection methods into one unified protocol, the invention reduces the number of separate steps, reagents, and operations required, thereby simplifying the overall methodology while preserving the ability to accurately identify multiple bacterial species simultaneously
3Reliability
If traditional culture methods are used, then bacterial identification can be performed, but the time required for results is too long for timely treatment
Solution Approach 1:
The patent replaces traditional mechanical culture methods with a molecular biology-based PCR system. Instead of relying on slow bacterial growth and manual culture techniques, the invention uses polymerase chain reaction to amplify and detect bacterial DNA directly from clinical samples, achieving rapid identification within hours rather than days while maintaining high reliability through specific primer and probe design
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 efficient identification of causative bacteria, facilitating quick treatment choices by allowing for the simultaneous examination of multiple bacterial species and their antimicrobial resistance genes.
Implementation Method 1
a step of performing a PCR method using a sample collected from a subject and a combination of sets of primers each specific to the full-length or a partial region of each of the gyrB gene of Escherichia coli, the nuc gene of Staphylococcus aureus, the atlE gene of Staphylococcus epidermidis, the gyrB gene of Klebsiella pneumoniae and the rpoB gene of Enterococcus spp.
Implementation Method 2
a step of detecting the presence or absence of amplification of the full-length or the partial region of each of the genes by using a combination of probes each specific to DNA of the full-length or the partial region
Data Source
AI summary
Disclosed is a method for identifying causative bacteria of sepsis, the method comprising: a step of performing a PCR method using a sample collected from a subject and a combination of sets of primers each specific to the full-length or a partial region of each of gyrB gene of Escherichia coli, nuc gene of Staphylococcus aureus, atlE gene of Staphylococcus epidermidis, gyrB gene of Klebsiella pneumoniae and rpoB gene of Enterococcus spp.; and a step of detecting the presence or absence of amplification of the full-length or the partial region of each of the genes by using a combination of probes each specific to DNA of the full-length or the partial region.