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Improvement of the Plaque Assay of Herpes Simplex Virus in HeLa Cells
Author(s) -
Yazaki Shigeyoshi,
Taniguchi Sadako,
Yoshino Kamesaburo
Publication year - 1966
Publication title -
japanese journal of microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0021-5139
DOI - 10.1111/j.1348-0421.1966.tb00301.x
Subject(s) - hela , titer , virus , herpes simplex virus , strain (injury) , virus quantification , virology , diluent , inoculation , chemistry , biology , cell culture , microbiology and biotechnology , in vitro , biochemistry , immunology , genetics , anatomy , nuclear chemistry
Plaque formation by the HF strain of herpes simplex virus in HeLa cells under agar overlay was possible when a line of this strain highly adapted to HeLa cells was used, whereas the homologous strain, passaged in eggs or lowly adapted to HeLa cells did not form any visible plaques using the same technique. Experiments were performed to investigate the influence of different diluents, cellular factors and conditions of virus adsorption. It was found that 0.1% yolk‐saline could replace the ordinarily used maintenance medium as a diluent. Among three lines of S3 HeLa cells maintained in different laboratories, a subclone possessing a rapid cellular growth rate showed the highest sensitivity to the virus. Monolayers which scarcely covered the whole glass surface were most suitable for virus inoculation. Such monolayers could be stored at 20 C for 2 days before use without lowering the efficiency of plaque formation. The highest titer of virus was obtained when adsorption of inoculated virus was allowed to take place at 37 C for 2 hr. This titer was about equal to the plaque titer obtained in primary chick embryo cell cultures.