PCR Primers and Probes for Foaming Bacteria Detection
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Solution Overview
Problem
Current methods for identifying and quantifying foaming and bulking bacteria in wastewater treatment processes are time-consuming, expensive, and require specialized equipment and expertise, making them impractical for routine use in wastewater treatment plants.
Innovation Solution
Development of PCR primers and probes specific to foaming-bulking bacteria genera and species, allowing for rapid and accurate detection and quantification using real-time PCR techniques, which can be performed with standard laboratory equipment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If light microscopy and FISH techniques are used to identify and quantify foaming-bulking bacteria, then measurement precision is improved, but device complexity and cost increase significantly
Solution Approach 1:
The patent replaces complex mechanical/optical microscopy systems with a biochemical PCR-based detection system. Instead of using light microscopy with FISH probes requiring expensive equipment and trained personnel, the invention uses PCR amplification with specific primers and probes that can be detected through simpler, more cost-effective means, thereby substituting a complex mechanical/optical system with a biochemical assay system.
Solution Approach 2:
The patent creates molecular copies (DNA amplification) of the target bacterial sequences through PCR. By amplifying specific DNA regions from foaming-bulking bacteria using genus-specific and species-specific primers, the method generates sufficient copies for detection without requiring direct visualization through complex microscopy systems, thus achieving accurate identification with simpler equipment.
2Measurement precision
If light microscopy and FISH techniques are used to identify and quantify foaming-bulking bacteria, then measurement precision is improved, but loss of time increases due to manual analysis requirements
Solution Approach 1:
The patent replaces time-consuming manual microscopy analysis with an automated biochemical PCR system. The PCR amplification process automatically multiplies target DNA sequences, and detection methods such as gel electrophoresis or real-time PCR provide results without requiring manual image analysis by trained personnel, thereby significantly reducing analysis time while maintaining or improving precision.
Solution Approach 2:
The patent performs preliminary DNA extraction and amplification of target sequences before detection. By pre-amplifying the bacterial DNA using PCR with specific primers, the method prepares sufficient target material in advance, eliminating the need for time-consuming manual counting and analysis under the microscope, thus accelerating the overall identification process.
3Measurement precision
If light microscopy and FISH techniques are used to identify and quantify foaming-bulking bacteria, then measurement precision is improved, but ease of operation deteriorates due to requirement for highly trained personnel
Solution Approach 1:
The patent replaces the need for highly trained personnel to perform microscopy and FISH analysis with a standardized biochemical PCR protocol. The PCR-based method uses well-established laboratory techniques that are more widely taught and easier to standardize, reducing the dependency on highly specialized training while maintaining detection accuracy through specific primer and probe designs.
Solution Approach 2:
The patent enables the detection system to perform identification automatically through the specificity of the PCR primers and probes. The method self-differentiates between bacterial genera and species through sequence-specific amplification, reducing the need for expert interpretation and manual analysis, thereby making the operation easier and more accessible to personnel with standard laboratory training.
4Measurement precision
If light microscopy and FISH techniques are used to identify and quantify foaming-bulking bacteria, then measurement precision is improved, but loss of time increases due to slow processing speed
Solution Approach 1:
The patent performs preliminary DNA extraction and PCR amplification of target sequences before final detection and quantification. By pre-amplifying the bacterial DNA using genus-specific and species-specific primers, the method prepares sufficient target material in advance, enabling rapid quantification without requiring time-consuming manual counting under the microscope, thus accelerating processing while maintaining accuracy.
Solution Approach 2:
The patent replaces slow manual microscopy quantification with automated PCR-based detection methods. Real-time PCR or post-PCR analysis techniques provide quantitative results through automated signal detection and analysis, significantly increasing processing speed compared to manual cell counting and analysis under the microscope.
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
Enables fast, inexpensive, and accurate assessment of foaming-bulking bacteria presence and abundance, facilitating better operational management and treatment interventions in wastewater treatment plants.
Implementation Method 1
performing a PCR reaction with such bacterial DNA in the presence of FBB-specific primers
Implementation Method 2
assaying for the presence of amplification products from the PCR reaction with genera-specific and species-specific probes
Data Source
AI summary
The present invention is directed to the detection of problematic foaming and bulking bacterial species in the biological wastewater treatment process. The invention provides various compositions of matter and methods for the detection of foaming and bulking bacterial species and genera in wastewater and other samples. PCR primers capable of amplifying 16s rRNA gene sequences from various foaming and bulking bacterial species are provided, as are probes that will specifically hybridize with PCR amplification products produced by the disclosed primers. In certain embodiments, the use of the disclosed PCR primers and probes in detection assays is disclosed.