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A quantitative assay for measuring human foamy virus using an established indicator cell line.
1Institute of Biotechnology in Medicine, School of Medical Technology and Engineering, National Yang-Ming University, 155 Section 2, Lie-Nong Street, 112, Taipei, Taiwan, ROC
Journal of Virological Methods
|May 5, 2001
Summary
A new indicator cell line, IdB14, enhances human foamy virus (HFV) detection. The foamy virus activated luciferase (FAL) assay offers a sensitive and rapid method for diagnosing and quantifying HFV infections.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Human foamy virus (HFV) is a complex retrovirus.
- Accurate and sensitive detection methods for HFV are crucial for diagnosis and research.
Purpose of the Study:
- To establish a novel indicator cell line for improved HFV detection.
- To develop a rapid and sensitive assay for HFV quantitation.
Main Methods:
- Co-transfection of baby hamster kidney-21 cells with plasmids containing a G418 resistance marker and the luc gene under an inducible HFV long terminal repeat (LTR) promoter.
- Selection and characterization of a stable subclone (IdB14) with low basal luciferase activity.
- Development of the foamy virus activated luciferase (FAL) assay.
Main Results:
- The IdB14 cell line demonstrated stable, low basal luciferase activity.
- The FAL assay showed time-dependent luciferase activity changes upon HFV infection, with differentiation possible by day 2.
- The FAL assay sensitivity was 400-fold higher than TCID(50) syncytium formation.
- HFV LTR promoter specificity and a linear relationship between multiplicity of infection (MOI) and luciferase expression were confirmed.
Conclusions:
- The IdB14 indicator cell line provides a stable platform for HFV detection.
- The FAL assay is a simple, rapid, and highly sensitive technique for diagnosing and quantifying active HFV infections.