A fast and visual method for duplex shrimp pathogens detection with high specificity using rapid PCR and molecular...

Area of Science:

• Molecular Biology
• Pathogen Detection
• Aquaculture Diagnostics

Background:

• Traditional Polymerase Chain Reaction (PCR) for pathogen detection is effective but time-consuming.
• Accurate and rapid diagnosis of shrimp pathogens is crucial for preventing disease outbreaks in aquaculture.
• Existing methods often require specialized equipment and trained personnel, limiting their accessibility.

Purpose of the Study:

• To develop a rapid, visual, and duplex molecular diagnostic method for simultaneous detection of two shrimp pathogens.
• To combine rapid PCR with molecular beacons for enhanced speed and specificity.
• To enable naked-eye determination of results for increased accessibility and reduced costs.

Main Methods:

• A duplex molecular assay utilizing rapid PCR technology was developed.
• Molecular beacons were employed for specific and simultaneous detection of target pathogen DNA.
• Automated equipment facilitated rapid thermal cycling, and a simple visual aid device was used for result interpretation.

Main Results:

• The developed method achieved specific detection of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Vibrio parahaemolyticus (VP) in a single reaction.
• Visual fluorescence changes allowed for naked-eye determination of results, eliminating the need for complex instrumentation.
• The entire detection process, from DNA extraction to result interpretation, was completed within 15 minutes.

Conclusions:

• This rapid, low-cost, and highly specific duplex visual detection method offers a significant advancement in shrimp pathogen diagnostics.
• The 15-minute turnaround time and visual readout facilitate rapid decision-making in disease prevention and management.
• This approach holds potential for widespread application in aquaculture settings, contributing to improved pathogen control strategies.