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Factors affecting virus plaque confirmation procedures.

D R Dahling1, G Sullivan, R W Freyberg

  • 1Virology Branch, Microbiology Research Division, Environmental Monitoring Systems Laboratory, Cincinnati, Ohio.

Journal of Virological Methods
|April 1, 1989
PubMed
Summary
This summary is machine-generated.

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Accurate virus quantification relies on direct plaque counts. The stab and scrape method for picking plaques offers the most reliable virus counts, minimizing false positives from transfer techniques.

Area of Science:

  • Virology
  • Cell Biology
  • Microbiology

Background:

  • Accurate virus quantification is crucial for virological studies.
  • The monolayer cell culture assay is a standard method for virus isolation and enumeration.
  • False-positive plaques can affect the reliability of virus counts.

Purpose of the Study:

  • To evaluate the reliability of direct plaque counts using the monolayer cell culture assay.
  • To compare different plaque-picking methods and storage conditions for virus samples.
  • To identify factors contributing to false-positive plaque rates.

Main Methods:

  • Direct plaque counts were performed using the monolayer cell culture assay.
  • Plaques were picked using the stab and scrape method and the stab only method.

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  • Plaque isolates were compared to laboratory stock viruses and sewage plaque isolates.
  • Storage conditions for picked plaques were evaluated (-70°C with or without fetal calf serum).

    • Main Results:

    • Direct plaque counts reliably confirmed the number of viruses isolated.
    • The stab and scrape method, followed by immediate transfer to cell culture tubes, yielded the most reliable counts.
    • Laboratory stock viruses showed higher confirmation rates than sewage plaque isolates.
    • A significant portion of unconfirmed plaques may result from plaque transfer techniques rather than true false positives.

    Conclusions:

    • The stab and scrape method is superior for obtaining reliable virus plaque counts.
    • Careful plaque transfer techniques are essential to minimize false-positive rates.
    • Virus type and sample source can influence plaque confirmation rates.