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High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
Permanent canine kidney (MDCK) cells for isolation and plaque assay of influenza B viruses
Abstract:
A wide range of influenza B virus strains with various passage histories uniformly formed well-defined clear plaques with high efficiency in cultures of an established line of canine kidney cells (MDCK). PFU titers of the viruses assayed in MDCK exceeded the titers assayed in ovo. With recently isolated strains such as B/Hong Kong/5/72 and Gifu/2/73, the PFU/EID50 ratios were as high as 100 to 400. MDCK cells have been successfully employed for primary isolation of influenza B viruses from throat washings of patients by direct plaquing.
Insights
Canine kidney cells (MDCK) efficiently plaque diverse influenza B virus strains, offering higher titers than traditional methods. This cell line enables direct isolation of influenza B from patient samples.
Area of Science:
- Virology
- Cell Biology
Background:
- Influenza B virus characterization and isolation are crucial for public health.
- Established cell lines are vital for efficient viral propagation and titration.
Purpose of the Study:
- To evaluate the efficacy of Madin-Darby Canine Kidney (MDCK) cells for influenza B virus plaque formation and isolation.
- To compare plaque-forming unit (PFU) titers in MDCK cells versus in ovo assays.
Main Methods:
- Culturing of Madin-Darby Canine Kidney (MDCK) cells.
- Inoculation with various influenza B virus strains.
- Plaque assays to determine viral titers.
- Comparison of PFU titers with egg infectious dose 50 (EID50) values.
- Testing MDCK cells for primary isolation from clinical samples.
Main Results:
- Influenza B virus strains uniformly formed well-defined plaques in MDCK cells with high efficiency.
- PFU titers in MDCK cells were consistently higher than in ovo titers.
- High PFU/EID50 ratios (100-400) were observed for recent strains.
- Successful primary isolation of influenza B virus from patient throat washings using direct plaquing in MDCK cells.
Conclusions:
- MDCK cells provide a sensitive and efficient system for influenza B virus plaque assays.
- MDCK cell cultures are superior to in ovo methods for quantifying influenza B virus infectivity.
- Direct plaquing in MDCK cells is a viable method for the primary isolation of influenza B virus from clinical specimens.
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