Multiplex PCR System for Pathogen Detection and Resistance Gene Identification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current diagnostic methods for sepsis, particularly those involving microarray and multiplex PCR, face challenges such as high costs, limited pathogen detection capabilities, inability to discriminate between gram-negative and gram-positive bacteria, and high false positive rates due to background contamination in blood culture bottles, leading to delayed antibiotic therapy and antibiotic resistance issues.
Innovation Solution
A method utilizing a single multiplex PCR system that differentiates between contaminating pathogens and infectious pathogens, eliminating the need for purification steps and allowing for simultaneous detection of gram-positive, gram-negative bacteria, and fungal infections, as well as identifying antimicrobial resistance genes, without requiring additional testing to distinguish between contamination and true infections.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If microarray and multiplex PCR methods are used for pathogen detection, then detection capability is improved, but device complexity and cost increase significantly
Solution Approach 1:
The patent combines multiple detection functions (bacterial detection, fungal detection, resistance gene detection) into a single multiplex PCR system that operates within a closed blood culture bottle, eliminating the need for separate purification and detection steps while maintaining comprehensive pathogen detection capability
Solution Approach 2:
The invention creates a universal detection system that can identify multiple types of pathogens (gram-positive bacteria, gram-negative bacteria, fungi) and their resistance genes simultaneously using a single reagent cartridge and amplification process, making the system adaptable to various detection needs without requiring separate specialized systems
2Measurement precision
If traditional culturing methods are used, then detection accuracy is maintained, but time consumption increases significantly
Solution Approach 1:
The system performs preliminary amplification of pathogen DNA directly within the blood culture bottle during the incubation period, so that when the culture becomes positive, the DNA is already amplified and ready for immediate detection, eliminating the need to wait for culture results before initiating molecular detection
Solution Approach 2:
The amplification process continues concurrently with the culture incubation process, maintaining continuous useful action rather than requiring sequential steps, thereby reducing total diagnosis time while preserving detection accuracy through the same molecular detection methods
3Reliability
If broad band antibiotic therapy is initiated early, then patient survival is improved, but antibiotic resistance development increases
Solution Approach 1:
The system provides rapid feedback on the specific pathogen identity and resistance genes present, enabling clinicians to adjust antibiotic therapy from broad-spectrum to targeted treatment based on actual infection findings, thereby improving outcomes while reducing resistance pressure from unnecessary broad-band coverage
4Measurement precision
If nested PCR with multiple primer sets is used, then detection sensitivity is improved, but false positive rate from contamination increases
Solution Approach 1:
The invention extracts and amplifies DNA directly from the blood culture bottle contents without requiring separate purification steps that could introduce contamination, and uses a single multiplex primer set that is optimized to reduce non-specific binding and false positives while maintaining sensitivity through direct amplification from the culture medium
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 approach significantly reduces false positives, enables timely antibiotic therapy, and improves patient outcomes by accurately identifying pathogens and resistance genes, thereby enhancing antimicrobial stewardship and reducing antibiotic resistance development.
Implementation Method 1
subjecting the sample to a single multiplex polymerase chain reaction (PCR), wherein said method comprises amplification of PCR products
Implementation Method 2
Electrochemical detection of bacterial and/or fungal infections
Data Source
AI summary
The present disclosure relates to methods and devices for amplifying a plurality of targets in a single PCR run while distinguishing between clinically relevant amplification and amplification from other sources such as from background contamination. The methods and devices further enable discrimination between gram-positive, gram-negative and fungal infections as wells as identify antimicrobial resistance genes. When applying the methods and devices of the invention, the species or genus of an infection(s), and genus of a fungal co-infection(s) or category of bacterial (gram-positive or negative) co-infection(s) are identified. Species identification of co-infections can also be achieved. Further, when applying the methods and devices of the invention, organisms which are likely to be contaminating organisms from a blood draw are identified.