CRISPR Primer Composition for Rapid L. acidophilus Strain Discrimination
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Solution Overview
Problem
Existing methods for discriminating and identifying Lactobacillus acidophilus strains are time-consuming, labor-intensive, and limited in detecting a wide range of strains, particularly due to mutations in specific genes and the need for complex genome data comparison.
Innovation Solution
A primer composition comprising specific forward and reverse primers (SEQ ID NOS: 1-16) targeting the CRISPR region for rapid and accurate discrimination and identification of Lactobacillus acidophilus species, strains, and the YT1 strain, using PCR for sample detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If 16S rRNA gene base sequence analysis is used for species discrimination, then identification can be performed, but the method is time-consuming and labor-intensive
Solution Approach 1:
The patent extracts and targets a specific CRISPR region within the bacterial genome for PCR amplification, rather than performing comprehensive whole-genome or 16S rRNA sequencing. This selective extraction of a diagnostic genomic region enables rapid species identification while maintaining accuracy, directly resolving the contradiction between identification precision and detection time.
Solution Approach 2:
The patent creates PCR copies of the CRISPR region through amplification, generating detectable DNA fragments that can be quickly analyzed to identify species. This copying approach replaces time-consuming sequencing methods with rapid PCR-based discrimination, achieving both speed and accuracy in species identification.
2Measurement precision
If RecA gene-based discrimination kit is used, then detection can be performed, but only 95% of strains can be detected due to mutations
Solution Approach 1:
The patent designs universal primers that target the CRISPR region, which is highly conserved across different Lactobacillus acidophilus strains. This universal primer set can detect all strains regardless of mutations in other genomic regions, achieving 100% strain detection coverage while maintaining high accuracy, thus resolving the contradiction between detection coverage and precision.
3Measurement precision
If NGS whole genome analysis is used for species discrimination, then comprehensive identification can be achieved, but the process is complicated and time-consuming
Solution Approach 1:
The patent extracts and focuses analysis on a specific CRISPR region rather than performing comprehensive whole-genome sequencing. This extraction approach maintains high identification accuracy for both species and strains while dramatically simplifying the analysis process and reducing time requirements, directly addressing the contradiction between precision and complexity.
4Measurement precision
If specific gene targeting methods are used, then detection can be performed, but many mutations can be made to specific genes limiting detection of various microorganisms
Solution Approach 1:
The patent targets the CRISPR region with universal primers that are highly conserved across all Lactobacillus acidophilus strains. This universal targeting approach maintains high detection accuracy while being adaptable to all strains regardless of mutations in other genes, resolving the contradiction between detection precision and versatility.
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 primer composition enables quick, precise discrimination and detection of Lactobacillus acidophilus strains, reducing time and effort, and is applicable in food, clinical, and environmental samples, supporting the development of functional food additives and health supplements.
Implementation Method 1
A primer composition comprising specific forward and reverse primers (SEQ ID NOS 1-8 and 9-16) for PCR, targeting the CRISPR region, allowing for rapid and accurate identification of Lactobacillus acidophilus species
Data Source
AI summary
The present disclosure relates to a composition for discriminating the species Lactobacillus acidophilus, which enables microorganisms belonging to the species Lactobacillus acidophilus to be discriminated, detected and identified simply, quickly and accurately from a target sample. Therefore, the present disclosure can be effectively used in the development of fermented milk, baby food, dairy products, livestock feed, cosmetics, health supplements, drugs for intestinal disorders, raw materials, etc. using the Lactobacillus acidophilus sp. strain.