Salmonella Binding Nucleic Acid Molecule for Specific Detection
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Solution Overview
Problem
Current methods for detecting Salmonella, particularly those belonging to Groups O4, O7, and O9, face challenges such as non-specific growth in selective media and complex primer design in gene amplification methods, and existing aptamers lack sufficient binding efficiency for practical use.
Innovation Solution
A novel nucleic acid molecule specifically binding to Salmonella, comprising polynucleotides with specific base sequences or modifications, is developed for enhanced detection, including those that can bind to Salmonella via deletion, substitution, insertion, or addition of bases, with a binding identity of 80% or more, and complementary sequences under stringent conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a selective medium is used to detect Salmonella, then Salmonella can be selectively grown, but Citrobacter and other opportunistic pathogens are also grown causing false positives
Solution Approach 1:
The detection process is segmented into multiple steps: first cultivating bacteria in a growth medium, then transferring to a selective medium for enrichment, and finally performing PCR amplification with specific primers. This multi-stage segmentation allows each step to address specific challenges - cultivation increases sensitivity, selective medium enriches Salmonella, and PCR provides specific identification, collectively resolving the contradiction between detection accuracy and reliability.
2Measurement precision
If gene amplification method (PCR) is used to specifically amplify Salmonella sequence, then specific detection is achieved, but primer design becomes complicated and time is required for amplification reaction
Solution Approach 1:
The selective medium enrichment step is performed as a preliminary action before PCR amplification. This pre-enrichment concentrates Salmonella cells in the sample, reducing the cycle threshold and allowing for faster PCR amplification with fewer cycles. The preliminary enrichment action thus reduces the overall detection time while maintaining high detection accuracy.
3Ease of operation
If existing aptamer is used for Salmonella detection, then detection method is simplified, but binding efficiency is insufficient for practical use
Solution Approach 1:
The aptamer sequence parameters are optimized by modifying the nucleotide composition, length, and structural characteristics. Specific changes in base sequence composition and secondary structure formation enhance the binding affinity and specificity to Salmonella surface antigens, thereby improving binding efficiency while maintaining the simplicity of the aptamer-based detection method.
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
The nucleic acid molecule effectively binds to Salmonella, allowing for specific detection of pathogenic Salmonella strains, improving detection accuracy and efficiency in food management and public health applications.
Implementation Method 1
a polynucleotide composed of a base sequence complementary to a polynucleotide which hybridizes to the polynucleotide (a) composed of any of the base sequences under stringent conditions and is bound to Salmonella
Data Source
AI summary
A nucleic acid molecule utilizable for Salmonella detection is provided. The nucleic acid molecule which binds to Salmonella includes any of the following polynucleotides (a) to (d): (a) a polynucleotide composed of any of base sequences of SEQ ID NOs: 1 to 17; (b) a polynucleotide composed of a base sequence obtained by deletion, substitution, insertion, and/or addition of one or more bases in any of the base sequences in the polynucleotide (a) and is bound to Salmonella; (c) a polynucleotide composed of a base sequence having an identity of 80% or more to any of the base sequences in the polynucleotide (a) and is bound to Salmonella; and (d) a polynucleotide composed of a base sequence complementary to a polynucleotide which hybridizes to the polynucleotide (a) composed of any of the base sequences under stringent conditions and is bound to Salmonella.