Universal Oligonucleotide Primers for Microbial Detection
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Solution Overview
Problem
Current primer sets for detecting microorganisms, particularly in clinical diagnostics, face challenges due to contamination and universality concerns, leading to decreased sensitivity and failure in diagnosing rare or unusual bacterial infections.
Innovation Solution
The use of specific oligonucleotide primers with sequences from SEQ ID NOs: 1-29, designed for bacterial, babesia, mycobacteria, and fungal detection, which are optimized for universal detection and amplification of conserved ribosomal DNA or RNA regions, enhancing sensitivity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If standardized published primer sets are used for microbial detection, then the detection process is simple and standardized, but the sensitivity decreases due to contamination and limited universality
Solution Approach 1:
The patent modifies primer parameters by adding 5'-extensions containing universal binding sites to historical primer sequences. This parameter change allows the primers to bind to a broader range of microbial targets while maintaining the simplicity of standardized PCR protocols. The extensions include universal sequences that enable amplification of diverse microorganisms without requiring new detection methodologies.
Solution Approach 2:
The patent creates universal primers that can detect multiple types of microorganisms (bacteria, fungi, parasites) simultaneously. The 5'-extensions contain universal binding sites that confer multi-functionality, allowing a single primer set to serve multiple detection purposes across different microbial taxa, thereby improving reliability without sacrificing operational simplicity.
2Ease of manufacture
If historical primer sets are used, then the primer design is simple and based on established protocols, but the detection fails for rare or unusual bacterial infections
Solution Approach 1:
The patent performs preliminary action by adding universal binding sites to primer sequences before the actual detection process. These 5'-extensions are designed in advance to accommodate a wide range of microbial targets, including rare and unusual pathogens. This preliminary modification ensures that the primers are pre-configured for broad detection capability while maintaining ease of manufacture through standard oligonucleotide synthesis.
3Reliability
If universal primers with extended sequences are used, then the detection sensitivity and specificity improve, but the primer sequence complexity increases
Solution Approach 1:
The patent segments the primer into two functional parts: the original historical primer sequence that provides specificity for particular microbial groups, and the added 5'-extension that provides universal binding capability. This segmentation allows each part to perform its specialized function while together they achieve improved sensitivity and specificity without excessive complexity.
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
These primers improve the detection sensitivity and specificity for bacteria, babesia, mycobacteria, and fungi, enabling accurate identification of microbial content in samples, even for rare or unusual pathogens.
Implementation Method 1
The use of specific oligonucleotide primers with sequences from SEQ ID NOs: 1-29, designed for bacterial, babesia, mycobacteria, and fungal detection, which are optimized for universal detection and amplification of conserved ribosomal DNA or RNA regions
Data Source
AI summary
Methods, compositions and kits for detection of a taxon of microorganisms in a sample are provided.