Development of a rapid visual detection technology for BmNPV based on CRISPR/Cas13a system

Area of Science:

• Molecular biology
• Virology
• Agricultural science

Background:

• Bombyx mori nucleopolyhedrovirus (BmNPV) causes significant losses in the sericulture industry due to frequent outbreaks and high mortality.
• Current detection methods for BmNPV lack sensitivity, specificity, and accessibility for on-site applications.
• There is a critical need for a rapid, accurate, and field-deployable diagnostic tool for BmNPV.

Purpose of the Study:

• To develop a novel, highly sensitive, and specific visualized detection technology for BmNPV.
• To create a cost-effective and user-friendly diagnostic tool applicable in various sericulture settings.
• To address the limitations of existing BmNPV detection methods.

Main Methods:

• Integration of Recombinase Polymerase Amplification (RPA) with the CRISPR/Cas13a system.
• Development of a detection platform utilizing microplate lateral flow test strips and a portable fluorescence detector.
• Validation using positive standard plasmids and infected silkworm tissue samples.

Main Results:

• The established CRISPR/Cas13a BmNPV detection technology achieved a sensitivity of 1 copy/μL for plasmids and 1 fg/μL for the BmNPV genome.
• Detection was completed within a rapid timeframe of 30-45 minutes.
• The technology demonstrated high specificity when tested against various pathogens in silkworm tissues.

Conclusions:

• The developed visualized nucleic acid detection technology for BmNPV is highly sensitive, specific, cost-effective, and convenient.
• This method is suitable for field detection and integration into silkworm egg quality monitoring systems.
• The technology provides a valuable tool for managing BmNPV in sericulture, improving disease control and reducing economic losses.