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A modified immunofluorescence assay for detection of Japanese encephalitis virus-infected cells

L Kabilan1

  • 1Center for Research in Medical Entomology, Madurai, Tamil Nadu, India. lalithakabilan@yahoo.co.in

Acta Virologica
|March 12, 2002
PubMed

Insights

A new immunofluorescence assay method detects Japanese encephalitis virus (JEV) in mosquito cells without damaging viral glycoprotein epitopes. This modified technique offers a stable, convenient alternative for JEV detection and antibody screening.

Area of Science:

  • Virology
  • Immunology
  • Entomology

Background:

  • Japanese encephalitis virus (JEV) detection typically uses indirect immunofluorescence (IF) assay.
  • Acetone fixation in standard IF assays can damage critical epitopes on JEV glycoprotein.

Purpose of the Study:

  • To develop a modified IF assay for identifying JEV-infected mosquito cells.
  • To overcome limitations of acetone fixation in standard JEV detection methods.

Main Methods:

  • Utilized paraformaldehyde fixation followed by detergent treatment of suspended cells.
  • Applied virus-specific antibodies for immunofluorescence staining.
  • Examined JEV-infected brain cells from Toxorhynchitis splendens mosquito larvae.

Main Results:

  • Successfully identified JEV-positive cells via intracellular immunofluorescence staining.
  • Demonstrated that the modified assay preserves epitope integrity.
  • Showcased the ability to store stained cell suspensions for up to 4 weeks.

Conclusions:

  • The modified IF assay provides a reliable alternative for JEV detection in cells.
  • This method is suitable for screening hybridoma cell lines for anti-JEV antibody production.
  • Offers enhanced convenience and epitope preservation compared to standard IF assays.

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