Visualization of infectious pancreatic necrosis virus (IPNV) particles labeled with fluorescent probes

Area of Science:

• Virology
• Microscopy techniques
• Biotechnology

Background:

• Infectious pancreatic necrosis virus (IPNV) poses a significant threat to aquaculture.
• Accurate quantification of viral particles is crucial for understanding infection dynamics and developing control strategies.

Purpose of the Study:

• To develop and validate a rapid and accurate fluorescence microscopy method for enumerating IPNV particles.
• To enable specific counting of IPNV in mixed viral populations and determine the physical particle to infectivity ratio.

Main Methods:

• Labeling of IPNV particles using SYBR Green I or a combination of monoclonal antibody and FITC-conjugated secondary antibody.
• Visualization and analysis of labeled viral particles using fluorescence microscopy.
• Comparison of IPNV particle size with fluorescent phage T4 virions.

Main Results:

• Labeled IPNV particles were successfully visualized and quantified.
• The method demonstrated the ability to differentiate and count IPNV particles based on their size.
• Accurate enumeration of IPNV particles was achieved on filters and cell surfaces, even in the presence of other virions.

Conclusions:

• Fluorescence microscopy provides a rapid and accurate method for quantifying IPNV particles.
• This technique facilitates specific enumeration of IPNV and calculation of the physical particle to infectivity ratio, aiding in disease management and research.