GBS Detection Kit Using Hybridization Probes and Amplification
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Solution Overview
Problem
Current diagnostic assays for detecting Group B Streptococcus (GBS) face challenges in sensitivity, precision, and speed, particularly in identifying low target copy levels, and require multiple biochemical and serological tests for confirmation.
Innovation Solution
A hybridization assay probe and amplification kit are developed, utilizing specific oligonucleotide sequences that include target-complementary and non-complementary sequences, with detectable labels, to enhance sensitivity and specificity, allowing for real-time amplification and detection of GBS nucleic acids in biological samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If traditional biochemical and serological tests are used for GBS identification, then the detection can be performed with simple equipment, but the detection speed is slow and multiple tests are required for confirmation
Solution Approach 1:
The patent combines nucleic acid amplification and detection into a single real-time assay system. The amplification kit includes primers, probes with detectable labels, and reaction conditions that enable simultaneous amplification and detection of GBS nucleic acids, eliminating the need for separate biochemical tests and reducing confirmation steps while maintaining simplicity through a unified protocol
Solution Approach 2:
The patent replaces traditional mechanical/biochemical identification methods (gram strain, catalase reaction, hemolytic activity, biochemical tests) with a molecular biology-based amplification and detection system. This substitution uses nucleic acid hybridization and amplification reactions that can be monitored in real-time, dramatically increasing detection speed while providing definitive identification in a single test
2Measurement precision
If DNA probe tests are used for GBS identification, then the identification accuracy is improved, but the sensitivity for low target copy levels is insufficient
Solution Approach 1:
The patent performs nucleic acid amplification before detection, generating numerous copies of the target sequence from limited initial material. This preliminary amplification step ensures that even low copy numbers of GBS nucleic acid are sufficiently amplified to be detected with high sensitivity, while the subsequent hybridization with specific probes maintains high identification accuracy
Solution Approach 2:
The patent changes the concentration parameter of the target nucleic acid through amplification, transforming low copy numbers into high copy numbers that are easily detectable. The amplification process increases the quantity of target sequence by several orders of magnitude, enabling detection of low target copy levels while maintaining the specificity and accuracy provided by probe hybridization
3Reliability
If multiple biochemical tests are performed for GBS confirmation, then the specificity is improved, but the time required for diagnosis increases
Solution Approach 1:
The patent combines multiple specificities into a single probe design that targets unique GBS nucleic acid sequences. The probe is designed to hybridize specifically to GBS DNA or RNA, providing definitive identification in one step rather than requiring multiple separate biochemical tests, thereby maintaining high specificity while dramatically reducing diagnostic time
Solution Approach 2:
The patent replaces multiple sequential biochemical tests (gram strain, catalase, hemolytic activity, biochemical tests, serological tests) with a single molecular amplification and detection assay. This substitution uses the specificity of nucleic acid hybridization to achieve definitive GBS identification in one test, eliminating the time required to perform and interpret multiple separate tests while maintaining or improving specificity
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 solution significantly improves the sensitivity, precision, and speed of GBS detection, enabling qualitative and quantitative measurements, and effectively identifies low target copy levels, simplifying the diagnostic process by combining amplification and detection in real-time.
Implementation Method 1
The hybridization assay probe uses nucleic acid hybridization for the qualitative detection of Group B Streptococcal DNA and RNA
Implementation Method 2
a kit for amplifying a Streptococcus agalactiae nucleic acid sequence that may be present in a biological sample
Data Source
AI summary
Compositions, methods and kits for detecting Group B streptococci. Particularly described are oligonucleotides that are useful as amplification primers and hybridization probes for detecting very low levels of Group B streptococci nucleic acids.