Microbial Biotechnology
  Marine Biotechnology
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 14 No. 1, Issue of January 15, 2011
© 2011 by Pontificia Universidad Católica de Valparaíso -- Chile Received August 3, 2010 / Accepted December 12, 2010
DOI: 10.2225/vol14-issue1-fulltext-11  

Infectious Pancreatic Necrosis Virus (IPNV) enumeration through epifluorescence microscopy: technical aspects

Maria Soledad Pavlov1 · Yoanna Eissler*1 · Juan Carlos Espinoza1 · David E. Garland2 · Juan Kuznar1

1Centro de Investigación y Gestión de Recursos Naturales, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso, Chile
2Facultad de Ciencias del Mar y de Recursos Naturales, Universidad de Valparaíso, Montemar, Viña del Mar, Chile

*Corresponding author: yoanna.eissler@uv.cl

Financial support: Innova Chile/CORFO, grant 05 CT6IPD-22 (2008-2011) and Universidad de Valparaíso, Chile, grant DIPUV 11/2007.

Keywords: digital image analysis, fluorescence microscope methods, immunofluorescence, Infectious Pancreatic Necrosis Virus (IPNV), SYBR-Green I.

Abstract   Full Text

A method for counting Infectious Pancreatic Necrosis Virus (IPNV) through epifluorescence microscopy was analyzed in detail. Image processing and statistic considerations are included. The particle size of viruses was compared in different experimental conditions such as the staining of the virus with SYBR-Green I or with antibodies for specific fluorescence labeling of viral proteins. The type of surface used as mounting support was assayed as well. The results indicated that the most suitable method involves the mounting of the viral-containing suspension on a membrane filter followed by the staining with a monoclonal antibody specific for a viral protein combined with a FITC (fluorescein isothiocyanate)-conjugated secondary antibody.

Supported by UNESCO / MIRCEN network