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. 1968 Sep;16(9):1331–1336. doi: 10.1128/am.16.9.1331-1336.1968

Rhinovirus Plaque Formation in WI-38 Cells with Methylcellulose Overlay

Timothy M Dolan 1,1, James D Fenters 1,2, Patricia A Fordyce 1,3, Jacob C Holper 1
PMCID: PMC547650  PMID: 4300170

Abstract

A sensitive, reliable plaque assay system is described for five rhinoviruses using freshly prepared methylcellulose overlay and human embryonic diploid cells. Circular plaques with irregular edges, 2 mm in size, were formed by rhinoviruses 1A, 2, 6, and 13 after 6 or 7 days of incubation. A fifth rhinovirus, 17, formed a 1- to 2-mm feather plaque after 14 days of incubation. Plaque counts of rhinoviruses 1A and 13 were not affected by varying the pH of the overlay from 6.9 to 7.5. Plaque sizes and plaque-forming unit values of high passage rhinoviruses 1A and 13 were equivalent when tested at 26, 31, or 36 C. The rhinoviruses tested were sensitive to incubation at 40 C or heating at 50 C. Enhancement of plaques was observed when Mg++ was incorporated into agar overlays, but enhancement did not occur when Mg++ was added to methylcellulose overlays.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cate T. R., Douglas R. G., Jr, Johnson K. M., Couch R. B., Knight V. Studies on the inability of rhinovirus to survive and replicate in the intestinal tract of volunteers. Proc Soc Exp Biol Med. 1967 Apr;124(4):1290–1295. doi: 10.3181/00379727-124-31990. [DOI] [PubMed] [Google Scholar]
  2. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dulbecco R. Production of Plaques in Monolayer Tissue Cultures by Single Particles of an Animal Virus. Proc Natl Acad Sci U S A. 1952 Aug;38(8):747–752. doi: 10.1073/pnas.38.8.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fenters J. D., Gillum S. S., Holper J. C., Marquis G. S. Serotypic relationships among rhinoviruses. Am J Epidemiol. 1966 Jul;84(1):10–20. doi: 10.1093/oxfordjournals.aje.a120614. [DOI] [PubMed] [Google Scholar]
  5. Fiala M., Kenny G. E. Enhancement of rhinovirus plaque formation in human heteroploid cell cultures by magnesium and calcium. J Bacteriol. 1966 Dec;92(6):1710–1715. doi: 10.1128/jb.92.6.1710-1715.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HOTCHIN J. E. Use of methyl cellulose gel as a substitute for agar in tissue-culture overlays. Nature. 1955 Feb 19;175(4451):352–352. doi: 10.1038/175352a0. [DOI] [PubMed] [Google Scholar]
  7. Hozinski V. I., Seibil V. B., Pantelyeva N. S., Mazurova S. M., Novikova E. A. The rct40 and T50 markers and the characteristics of some variants of measles virus. Acta Virol. 1966 Jan;10(1):20–27. [PubMed] [Google Scholar]
  8. KISCH A. L., JOHNSON K. M. A plaque assay for respiratory syncytial virus. Proc Soc Exp Biol Med. 1963 Mar;112:583–589. doi: 10.3181/00379727-112-28111. [DOI] [PubMed] [Google Scholar]
  9. KISCH A. L., WEBB P. A., JOHNSON K. M. FURTHER PROPERTIES OF FIVE NEWLY RECOGNIZED PICORNAVIRUSES (RHINOVIRUSES). Am J Hyg. 1964 Mar;79:125–133. doi: 10.1093/oxfordjournals.aje.a120368. [DOI] [PubMed] [Google Scholar]
  10. LWOFF A., LWOFF M. L'inhibition du développement du virus poliomyélitique à 39 degrees et le problème du rôle de l'hyperthermie dans l'évolution des infections virales. C R Hebd Seances Acad Sci. 1958 Jan 6;246(1):190–192. [PubMed] [Google Scholar]
  11. PARSONS R., TYRRELL D. A. A plaque method for assaying some viruses isolated from common colds. Nature. 1961 Feb 25;189:640–642. doi: 10.1038/189640a0. [DOI] [PubMed] [Google Scholar]
  12. PORTERFIELD J. S. Titration of some common cold viruses (rhinoviruses) and their antisera by a plaque method. Nature. 1962 Jun 16;194:1044–1047. doi: 10.1038/1941044a0. [DOI] [PubMed] [Google Scholar]
  13. SCHULZE I. T., SCHLESINGER R. W. Plaque assay of dengue and other group B arthropod-borne viruses under methyl cellulose overlay media. Virology. 1963 Jan;19:40–48. doi: 10.1016/0042-6822(63)90022-9. [DOI] [PubMed] [Google Scholar]
  14. VOGT M., DULBECCO R., WENNER H. A. Mutants of poliomyelitis viruses with reduced efficiency of plating in acid medium and reduced neuropathogenicity. Virology. 1957 Aug;4(1):141–155. doi: 10.1016/0042-6822(57)90050-8. [DOI] [PubMed] [Google Scholar]
  15. WALLIS C., MELNICK J. L., BIANCHI M. Factors influencing enterovirus and reovirus growth and plaque formation. Tex Rep Biol Med. 1962;20:693–702. [PubMed] [Google Scholar]
  16. WEBB P. A., JOHNSON K. M., MUFSON M. A. A DESCRIPTION OF TWO NEWLY-RECOGNIZED RHINOVIRUSES OF HUMAN ORIGIN. Proc Soc Exp Biol Med. 1964 Jul;116:845–852. doi: 10.3181/00379727-116-29389. [DOI] [PubMed] [Google Scholar]

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