A rapid method for titration of ascovirus infectivity

Area of Science:

• Virology
• Insect Pathology
• Molecular Biology

Background:

• Ascoviruses represent a recently identified viral family.
• Traditional titration methods like plaque assays and end-point PCR are time-consuming and lack accuracy for ascoviruses.

Purpose of the Study:

• To develop a rapid, accurate, and simple method for titrating ascovirus stocks.
• To utilize ascovirus-induced apoptosis in insect cells as a basis for viral titration.

Main Methods:

• Insect cells were infected with serial dilutions of ascovirus (10^-2 to 10^-10).
• Cells were stained with trypan blue 24 hours post-infection to assess viability.
• The Reed and Muench method was adapted to calculate virus titer based on the percentage of positive wells, defined by stained cell rates exceeding control averages.

Main Results:

• The developed method provides a rapid and reproducible assessment of ascovirus titers.
• This apoptosis-based assay is less material-consuming compared to conventional titration techniques.
• The method eliminates the subjectivity inherent in other ascovirus titration procedures.

Conclusions:

• A novel, apoptosis-based assay offers a superior alternative for ascovirus titration.
• This method is accurate, simple, and efficient, addressing the limitations of existing techniques.
• The developed assay facilitates more reliable quantification of ascovirus stocks.