Multiplex PCR Kit for Phage Detection in Dairy Fermentation
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Solution Overview
Problem
Current methods for detecting Streptococcus thermophilus infecting phages in industrial dairy settings are inadequate, as they fail to efficiently identify the rapidly diversifying phage groups, particularly the 987 and 5093 groups, which can lead to fermentation spoilage and production issues.
Innovation Solution
A kit comprising specific primer pairs for PCR analysis that can simultaneously detect four distinct phage groups (987, 5093, cos-containing, and pac-containing phages) by generating amplicons of different sizes, allowing for rapid identification and differentiation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional PCR methods with single primer pairs are used, then detection of specific phage groups is achieved, but detection of all four phage groups (987, 5093, cos-containing, and pac-containing) cannot be performed simultaneously
Solution Approach 1:
The patent combines multiple primer pairs (first primer pair for 987 phages, second primer pair for 5093 phages, third primer pair for cos-containing phages, and fourth primer pair for pac-containing phages) into a single multiplex PCR reaction system. This allows simultaneous detection of all four phage groups in one assay, resolving the contradiction between detection versatility and method complexity by merging multiple detection capabilities into a unified protocol.
Solution Approach 2:
The invention creates a universal detection kit that can identify all four major phage groups infecting Streptococcus thermophilus using a single multiplex PCR methodology. The kit is designed to be universally applicable across different dairy fermentation contexts, eliminating the need for separate assays for each phage group and providing comprehensive phage surveillance capability.
2Measurement precision
If multiple separate PCR assays are performed for each phage group, then specific detection accuracy is maintained, but detection time and productivity are reduced
Solution Approach 1:
The patent merges four separate PCR detection protocols into a single multiplex PCR assay that simultaneously amplifies and detects all four phage groups. By performing all detections in parallel within one reaction tube using phage-group-specific primer pairs, the method maintains identification accuracy while reducing total detection time from sequential multiple assays to a single unified assay.
Solution Approach 2:
The multiplex PCR system enables continuous simultaneous detection of all four phage groups in a single uninterrupted reaction process. All primer pairs amplify their respective targets concurrently during the same thermal cycling protocol, eliminating the need to stop and restart between different assays, thereby maximizing detection productivity without sacrificing precision.
3Reliability
If phage detection is delayed, then fermentation process continues, but phage infection spreads and causes spoilage
Solution Approach 1:
The multiplex PCR detection method enables preliminary identification of phage infections at the earliest stages of contamination. By providing rapid simultaneous detection of all four phage groups, the system allows producers to take corrective actions (such as adjusting sanitation procedures or rotating starter cultures) before phage infection spreads throughout the fermentation process, thereby protecting fermentation stability.
Solution Approach 2:
The invention provides rapid feedback on phage presence across all four phage groups through simultaneous detection. This immediate feedback mechanism allows producers to quickly assess fermentation risk and implement appropriate countermeasures, creating a closed-loop system where detection results directly inform process control decisions to prevent fermentation spoilage.
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 efficient detection of all four phage groups, facilitating quicker response times and the selection of resistant Streptococcus thermophilus strains for starter culture production, thereby improving fermentation stability and product consistency.
Implementation Method 1
A kit comprising specific primer pairs for PCR analysis that can simultaneously detect four distinct phage groups (987, 5093, cos-containing, and pac-containing phages) by generating amplicons of different sizes
Data Source
AI summary
The present invention relates to a kit suitable for detection of Streptococcus thermophilus infecting phages, said kit comprising: (a) a first primer pair suitable to generate an amplicon characteristic for a 987 phage; and (b) a second primer pair suitable to generate an amplicon characteristic for a 5093 phage. Further, the present invention relates to a method for detecting the presence of Streptococcus thermophilus infecting phages in a sample.