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Updated: Aug 2, 2026
Method for Measurement of Viral Fusion Kinetics at the Single Particle Level
Published on: September 7, 2009
[Quantitative analysis of cell fusion caused by glycoprotein of paramyxoviruses with reporter gene method]
Objective:
To establish a kind of quantitative assay method for analyzing the reporter gene assay for fusion was established.
Methods:
Two populations of BHK21 cells were infected with vTF-7 recombinant and wild-type vaccinia viruses respectively and transfected with paramyxovirus F and HN cDNA and plasmid pG1NT7 beta-gal respectively. After 16 h, the two cell populations were removed from the wells by trypsinization. Equal numbers of the two populations were mixed in wells of a 96-well flat-bottom plate after washing and pelleting. After 15 hours' incubation at 37 degrees C, the cells were lysed with Nonidet P-40 and A values were read at 570 nm with an ELISA reader supported by SOFTMAX software.
Results:
Reporter gene method had a close relationship with cell count method, r = 0.9890(P < 0.01). Positive and negative coincidence rates were 100%. The best main reaction conditions are as follows: substrate concentration, 16 mmol/L; color reaction time, 20-25 min; amount of DNA for transfection, 1 microgram; cell numbers for each population, 1 x 10(5).
Conclusion:
Reporter gene method is a kind of very sensitive, specific and repeatable quantitative analysis for cell fusion and can be used in the studies on membrane fusion caused by any viruses.
