Multiplex PCR Antibiotic Susceptibility Detection Using RNA Response
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Solution Overview
Problem
Current methods for diagnosing infectious diseases using PCR are slow and impractical due to the need for multiple assays and low levels of pathogen nucleic acid, while antibiotic resistance detection requires lengthy bacterial growth processes, delaying timely treatment.
Innovation Solution
A method involving multiplex PCR with integrated sample preparation and detection systems, combined with RNA analysis to determine antibiotic susceptibility by incubating samples with multiple antibiotics, reverse-transcribing RNA, and quantifying gene expression patterns to assess resistance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple separate PCR assays are run for different pathogens, then diagnostic accuracy is improved, but diagnostic time and complexity increase
Solution Approach 1:
The patent combines multiple PCR assays targeting different pathogens into a single multiplex PCR reaction. Multiple primer sets specific to different pathogens are included in the same reaction mixture, allowing simultaneous amplification and detection of multiple pathogens from a single sample, thereby maintaining diagnostic accuracy while reducing time and complexity
Solution Approach 2:
The invention creates a universal diagnostic platform that can detect multiple types of pathogens (bacteria, viruses, fungi) using a single PCR system. The multiplex PCR assay serves multiple diagnostic functions simultaneously, eliminating the need for separate assays for each pathogen type
2Adaptability or versatility
If large panels of PCR assays are run for all possible causative organisms, then diagnostic coverage is improved, but resource requirements and cost increase
Solution Approach 1:
The patent merges multiple individual PCR assays into a single multiplex reaction, allowing detection of numerous pathogens simultaneously from one sample. This consolidation maintains comprehensive diagnostic coverage while using a single sample volume rather than requiring separate samples for each assay
Solution Approach 2:
The multiplex PCR panel includes primers for more pathogens than are likely to be present in any given sample, providing excessive coverage. This ensures that all possible causative organisms can be detected in a single assay, maintaining comprehensive diagnostic coverage while efficiently using sample volume
3Reliability
If nested secondary PCR reactions are performed to increase robustness, then detection sensitivity is improved, but risk of contamination and operational complexity increase
Solution Approach 1:
The patent combines multiple PCR amplifications into a single multiplex reaction rather than performing sequential nested PCRs. This single-step approach achieves robust detection of multiple pathogens simultaneously while eliminating the operational complexity and contamination risks associated with multiple handling steps
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 and accurate determination of antibiotic susceptibility for multiple bacteria and antibiotics, reducing diagnostic time and improving treatment efficacy.
Implementation Method 1
In recent years, the polymerase chain reaction (PCR) has become a method of choice for rapid diagnosis of infectious agents
Implementation Method 2
reverse-transcribing RNA for a plurality of genes that each show a different pattern of expression
Data Source
AI summary
Methods, sample vessels, and instruments are provided for determining susceptibility of a bacterium to one or more classes of antibiotics.