Novosphingobium Detection Primers for Activated Sludge
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Solution Overview
Problem
Current methods fail to specifically detect and quantify bacteria of the genus Novosphingobium in activated sludge, which is crucial for effective oil decomposition in drainage treatment, due to the presence of multiple microorganism species.
Innovation Solution
A method involving specific primers for amplifying and detecting a nucleotide sequence of the 16S rRNA gene, specifically targeting a region of the 16S rRNA gene of Novosphingobium, allowing for the specific detection and quantification of Novosphingobium bacteria even in mixed microbial environments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If predetermined primers for detecting Novosphingobium bacteria are used, then detection capability is provided, but specific detection in mixed microorganism systems cannot be achieved
Solution Approach 1:
The patent applies local quality by designing primers that specifically target a unique local region (nucleotides 540-675) of the 16S rRNA gene in Novosphingobium bacteria. This localized targeting ensures that only the specific bacterial genus is detected while ignoring other microorganisms in the mixed community, thereby achieving both detection specificity and reliability
Solution Approach 2:
The patent employs parameter changes by optimizing primer sequences to match the specific nucleotide sequence parameters of the target region in Novosphingobium 16S rRNA gene. By carefully selecting and adjusting the primer sequences (SEQ ID NO: 6 and SEQ ID NO: 7) to complement the target region parameters, the method achieves specific amplification and detection of Novosphingobium bacteria even in complex microbial environments
2Quantity of substance
If general detection methods are used in activated sludge, then microorganisms can be detected, but quantification of specific Novosphingobium bacteria is insufficient
Solution Approach 1:
The patent applies the extraction principle by isolating and amplifying only the specific nucleotide sequence region (540-675) of the 16S rRNA gene from Novosphingobium bacteria using specially designed primers. This extraction of the target sequence from the complex genomic material enables accurate quantification of Novosphingobium bacteria without interference from other microorganisms in the activated sludge
Solution Approach 2:
The patent uses PCR amplification as an intermediary process that specifically amplifies the target Novosphingobium DNA sequence between the primer binding sites. This intermediary amplification step creates sufficient quantity of the specific target sequence for accurate detection and quantification, while the primer specificity ensures only Novosphingobium DNA is amplified
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 precise detection and quantification of Novosphingobium bacteria, facilitating better control over oil removal reactions in drainage treatment by distinguishing them from other microorganisms.
Implementation Method 1
a first step of amplifying nucleotides using a test DNA as a template and primers capable of amplifying a nucleotide sequence of consecutive 94 or more nucleotides and 136 or less nucleotides in the nucleotide sequence of SEQ ID NO: 1 to obtain an amplified product
Data Source
AI summary
A method for detecting a bacterium of the genus Novosphingobium, includes a first step of amplifying nucleotides using a test DNA as a template and primers capable of amplifying a nucleotide sequence of consecutive 94 or more nucleotides and 136 or less nucleotides in the nucleotide sequence of SEQ ID NO: 1 to obtain an amplified product; and a second step of detecting the amplified product.