DNA Chip Identifying Filamentous Microorganisms via Segmented Monoplex PCR
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for identifying filamentous microorganisms causing sludge bulking in wastewater treatment are inefficient due to the complexity of bacterial flora and the limitations of conventional identification techniques, such as multiplex PCR, which suffer from decreased sensitivity and increased complexity and cost.
Innovation Solution
A DNA chip using oligonucleotide probes designed from the 16S-23S ribosomal DNA intergenic spacer sequences is employed for identifying filamentous microorganisms, allowing for the amplification and hybridization of target sequences to accurately identify specific microorganisms, including Leucothrix mucor, Leptothrix sp., Haliscomenobacter hydrossis, Nocardia sp., and others, using universal primers and PCR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiplex PCR is used to identify filamentous microorganisms, then identification capability is improved, but PCR sensitivity decreases
Solution Approach 1:
The patent divides the identification task into separate monoplex PCR reactions for different filamentous microorganism groups, each with its own specific primers. This segmentation allows each PCR reaction to focus on a specific target, maintaining high sensitivity while collectively covering multiple microorganism types through systematic grouping.
2Adaptability or versatility
If serial PCRs are used to identify different microorganisms, then identification capability is improved, but assay complexity increases
Solution Approach 1:
The patent segments the identification process into parallel monoplex PCRs performed simultaneously rather than sequentially. Multiple PCR reactions are conducted in parallel using different primer sets, reducing the time required compared to serial PCRs while maintaining the ability to identify multiple microorganism types.
Solution Approach 2:
The patent transitions from temporal sequencing (serial PCRs performed one after another) to spatial parallelization (multiple PCRs performed simultaneously in different reaction vessels). This dimensional change from time-based to space-based execution reduces overall assay complexity and time requirements.
3Adaptability or versatility
If multiplex PCR is used for filamentous microorganism identification, then identification capability is improved, but cost increases
Solution Approach 1:
The patent uses simple monoplex PCR reactions with single primer sets for each microorganism group rather than complex multiplex PCR requiring multiple primer sets. This segmentation simplifies the reagent requirements and reduces the cost of each individual reaction, making the overall identification process more economical.
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 DNA chip demonstrates 100% sensitivity and specificity in identifying target filamentous microorganisms, reducing cross-reactions and improving the efficiency of wastewater treatment by enhancing the ability to control sludge bulking.
Implementation Method 1
a DNA chip having a plurality of probes, wherein the probe comprises an oligonucleotide sequence... the 16S-23S ribosomal DNA intergenic spacer region sequence amplified in step (b) hybridize with the DNA chip
Data Source
AI summary
A DNA chip for identifying filamentous microorganisms, including a substrate and a plurality of probes, wherein the probe includes SEQ ID No. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, complementary sequences thereof, derivatives thereof or combinations thereof. The derivative is 5′ and/or 3′ end of the sequence SEQ ID No. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 with at least one thymidine residue added or is 5′ and/or 3′ end of the sequence SEQ ID No. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 with one or two nucleotides added or deleted.