New Detection Methods for Cryphonectria Hypovirus 1 (CHV1) through SYBR Green-Based Real-Time PCR and Loop-Mediated...

Area of Science:

• Plant Pathology
• Virology
• Biotechnology

Background:

• Mycoviruses can act as biocontrol agents, reducing phytopathogenic fungi impact and enhancing plant health.
• Cryphonectria hypovirus 1 (CHV1) significantly reduces the virulence of *Cryphonectria parasitica*, the chestnut blight pathogen.

Purpose of the Study:

• To introduce two novel diagnostic protocols for the accurate and efficient detection of CHV1.
• To evaluate the sensitivity and performance of these new assays compared to existing methods.

Main Methods:

• Development of a colorimetric loop-mediated isothermal amplification (LAMP) assay targeting CHV1 ORF A.
• Development of a real-time quantitative PCR (qPCR) assay targeting CHV1 ORF B.
• Comparative analysis of LAMP and qPCR against reverse transcription-PCR (RT-PCR) and the Bavendamm test.

Main Results:

• The LAMP assay demonstrated a tenfold increase in sensitivity compared to RT-PCR, with a visible color change within 35 minutes.
• The qPCR assay showed a remarkable 100-fold improvement in sensitivity over RT-PCR.
• Both LAMP and qPCR outperformed the Bavendamm test in distinguishing hypovirulent strains.

Conclusions:

• Two pioneering, highly sensitive diagnostic assays for CHV1 detection have been developed.
• These advanced methodologies represent a significant improvement for CHV1 surveillance.
• The assays hold great promise for advancing research in the biological control of *C. parasitica*.