Nucleic Acid Detection for Rapid Antibiotic Susceptibility Testing
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Solution Overview
Problem
Current methods for determining antibiotic susceptibility, particularly for slow-growing microorganisms like Neisseria gonorrhoeae, face challenges in achieving rapid and accurate detection due to their slow growth and lack of a classic transcriptional SOS response to DNA damage.
Innovation Solution
A method involving the quantitative detection of nucleic acids in antibiotic-treated samples, either with or without lysis treatment, to assess antibiotic susceptibility by comparing nucleic acid concentration ratios, allowing for the differentiation between susceptible and resistant microorganisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional antibiotic susceptibility testing methods are used for slow-growing microorganisms, then accuracy can be maintained, but the testing time becomes excessively long
Solution Approach 1:
The patent extracts the nucleic acid from the microorganism and detects it separately using PCR amplification. By taking out the nucleic acid detection from the traditional culture-based testing, the method achieves rapid results (within hours) without requiring slow microbial growth, thus resolving the contradiction between accuracy and testing time
Solution Approach 2:
The patent replaces the mechanical/biological process of microbial culture and growth with a molecular biology approach (PCR amplification of nucleic acid). This substitution eliminates the time-consuming growth phase while maintaining detection accuracy, directly addressing the time-accuracy contradiction
2Loss of time
If rapid detection methods are implemented for antibiotic susceptibility, then testing time is reduced, but detection accuracy and reliability deteriorate
Solution Approach 1:
The patent performs preliminary extraction of nucleic acid from the microorganism before antibiotic treatment. This preliminary action allows the detection process to begin independently of microbial growth, enabling rapid testing while maintaining reliability through direct molecular detection of the target organism
Solution Approach 2:
The patent uses PCR amplification with specific primers that provide feedback on the presence and quantity of target nucleic acid. This molecular feedback mechanism ensures reliable detection even in rapid testing conditions, as the amplification process provides clear signal amplification that can be reliably measured
3Measurement precision
If conventional culture-based methods are used for microorganisms lacking SOS response, then traditional susceptibility markers can be detected, but the methods fail to provide rapid results
Solution Approach 1:
The patent changes the detection parameter from phenotypic markers (which require gene expression and growth) to genotypic markers (nucleic acid sequences). This parameter change allows detection of susceptibility markers directly at the DNA/RNA level, enabling both precise measurement and high productivity simultaneously
Solution Approach 2:
The patent substitutes the biological mechanism of SOS response and gene expression with a direct nucleic acid amplification mechanism. This substitution bypasses the need for microbial growth and transcriptional responses, enabling rapid and high-throughput testing while maintaining detection precision for microorganisms that lack classic SOS responses
Data Source
AI summary
Provided herein is an antibiotic susceptibility test and related compositions, methods and systems based on detection of a nucleic acid from a target microorganism in a sample in the presence or absence of a lysis treatment of the sample.