Alpha-glucosidase Substrate for Streptococcus agalactiae Detection
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Solution Overview
Problem
Current methods for detecting and identifying Streptococcus agalactiae require multiple steps and supplementary tests, including selective media and biochemical assays, which can be cumbersome and lack direct enzymatic substrate-based detection, making them inefficient for rapid identification.
Innovation Solution
The use of α-glucosidase enzymatic substrates, such as indoxyl-derivative-based substrates, in a reaction medium for specific detection and identification of Streptococcus agalactiae, allowing for visualization of colonies without diffusion of coloration and without harming the bacteria, which can be used in both culture and visualization media.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If selective media and supplementary tests are used for detecting Streptococcus agalactiae, then detection reliability is improved, but detection time and complexity increase
Solution Approach 1:
The patent combines multiple detection functions into a single culture medium by incorporating both selective antibiotics (gentamicin, nalidixic acid, polymyxin) and the specific enzymatic substrate (5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside) that reacts with Streptococcus agalactiae β-glucosidase. This merging eliminates the need for separate selective media and supplementary biochemical tests, achieving reliable detection in a single step while reducing detection time.
Solution Approach 2:
The culture medium serves multiple functions simultaneously: it acts as a selective medium (inhibiting non-target bacteria), a differential medium (distinguishing Streptococcus agalactiae through color change), and a growth medium (supporting bacterial proliferation). This multi-functionality resolves the contradiction by providing reliable detection without requiring multiple separate tests.
2Productivity
If Granada medium is used for direct isolation and identification, then detection speed is improved, but operational complexity and equipment requirements increase due to anaerobic conditions
Solution Approach 1:
The patent changes the oxygen requirement parameter from anaerobic (Granada medium) to aerobic conditions. The culture medium contains antibiotics and enzymatic substrate that function effectively under aerobic conditions, allowing Streptococcus agalactiae to be detected through color change without requiring specialized anaerobic incubation equipment or procedures, thus improving ease of operation while maintaining detection speed.
3Measurement precision
If multiple supplementary tests are performed for accurate identification, then measurement precision is improved, but device complexity and procedure complexity increase
Solution Approach 1:
The patent uses a chromogenic substrate (5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside) that produces a distinct color change (formation of 5,5'-dibromo-4,4'-dichloro-indigo) when cleaved by Streptococcus agalactiae β-glucosidase. This colorimetric reaction provides precise identification of the bacterium directly on the culture medium, eliminating the need for multiple supplementary biochemical tests and reducing procedure complexity while maintaining high identification precision.
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
This method enables rapid and specific detection of Streptococcus agalactiae with high sensitivity and specificity, reducing the need for multiple tests and improving the efficiency of identification, particularly in clinical samples, by using α-glucosidase enzymatic substrates that produce a concentrated color change at the colonies.
Implementation Method 1
The medium contains 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside, which is a substrate for the enzyme α-glucosidase, present in Streptococcus agalactiae. The medium consequently changes color from blue to green as a result of the hydrolysis of the substrate by the enzyme.
Data Source
AI summary
The present invention concerns a method for specifically detecting and identifying Streptococcus agalactiae, using a reaction medium comprising at least one α-glucosidase enzymatic substrate.