Multiplex PCR Assays for Specific Shrimp Pathogen Detection
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Solution Overview
Problem
Current diagnostic methods for detecting Hepatobacter penaei (NHP) and Vibrio parahaemolyticus (AHPND) in shrimp are limited, particularly when non-specific amplifications occur, and there is a need for alternative PCR and real-time PCR assays to accurately detect and confirm the presence of these pathogens.
Innovation Solution
Development of PCR-based assays that can detect specific genes such as flgE from H. penaei, shrimp 18s rRNA, bacterial 16S rRNA, Vibrio pirA, and Vibrio pirB genes, which can be used in single or multiplex reactions to identify multiple pathogens simultaneously.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional PCR methods are used to detect H. penaei and V. parahaemolyticus, then diagnostic capability is provided, but non-specific amplifications occur reducing detection accuracy
Solution Approach 1:
The patent divides the detection process into multiple specific PCR assays, each targeting distinct gene regions of H. penaei and V. parahaemolyticus. By segmenting the detection into separate specific assays rather than using a single conventional PCR method, the patent achieves reliable detection while avoiding non-specific amplification through targeted primer design for specific pathogen genes.
Solution Approach 2:
The patent applies local quality by designing primers that specifically target unique gene sequences of each pathogen (H. penaei and V. parahaemolyticus). This localized specificity ensures that each PCR assay detects only its intended target, improving both reliability and precision by eliminating cross-reactivity and non-specific amplification in the detection system.
2Adaptability or versatility
If multiple diagnostic methods are developed for different pathogens, then detection capability increases, but diagnostic system complexity increases
Solution Approach 1:
The patent creates a universal PCR-based diagnostic platform that can detect multiple pathogens (H. penaei and V. parahaemolyticus) using standardized protocols and reagents. This multi-functional approach allows the same basic PCR methodology to be applied across different pathogen detections, increasing versatility while maintaining system simplicity through methodological consistency.
Solution Approach 2:
The patent segments the diagnostic capability into modular, pathogen-specific PCR assays that can be independently performed or combined. Each assay targets a specific pathogen with dedicated primers, allowing the system to scale versatility by adding or removing specific assays without increasing overall system complexity, as each module operates independently with standardized procedures.
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 assays provide accurate and efficient detection of H. penaei and V. parahaemolyticus, enabling effective treatment and reducing the impact of these diseases in shrimp farming by identifying infected populations.
Implementation Method 1
various PCR based assays that can detect H. penaei and/or Vibrio parahaemolyticus (V. parahaemolyticus) that causes AHPND in a nucleic acid sample obtained from one or more shrimp
Data Source
AI summary
Described herein various PCR based assays that can detect H. penaei and/or V. parahaemolyticus that causes AHPND in a nucleic acid sample obtained from one or more shrimp. In some aspects, the PCR based assays can detect one or more of the following genes: the flgE gene from H. penaei, the shrimp 18s rRNA gene, the shrimp beta actin gene from shrimp, a bacterial 16S rRNA gene from bacteria (any type of bacteria) the Vibrio pirA gene, and the Vibrio pirB gene. The assays described herein can be single assays or can be multiplexed such that more than one gene and/or more than one bacterial species can be detected in a single reaction. Other compositions, compounds, methods, features, and advantages of the present disclosure will be or become apparent to one having ordinary skill in the art upon examination of the following drawings, detailed description, and examples.