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Given biosafety concerns in virus research, the use of viral substitutes, such as bacteriophages, has been well accepted, presenting results comparable to those of the target viruses under the same experimental conditions. The double-layer agar plate (DLAP) protocol is based on the ability of bacteriophages to infect and lyse growing bacterial cells. In this method is necessary to use two layers of agar, where the first is the base layer and the second is a softer agar layer made of agar and a mix of host bacteria. DLAP protocol is a widely used, low-cost method for the enumeration and detection of bacteriophages in food, water, and beverages. In this chapter, we used the Phi6 SARS-CoV-2 surrogate as an example.

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Double-Layer Agar Plaque Protocol for Propagation and Quantification of SARS-CoV-2 Surrogate, Bacteriophage Phi6: Techniques and Applications

  • 章节

  • 发表日期：

    2026年

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  • Ruthchelly Tavares da Silva,
  • Edson Douglas Silva Pontes,
  • Marciane Magnani

摘要

Given biosafety concerns in virus research, the use of viral substitutes, such as bacteriophages, has been well accepted, presenting results comparable to those of the target viruses under the same experimental conditions. The double-layer agar plate (DLAP) protocol is based on the ability of bacteriophages to infect and lyse growing bacterial cells. In this method is necessary to use two layers of agar, where the first is the base layer and the second is a softer agar layer made of agar and a mix of host bacteria. DLAP protocol is a widely used, low-cost method for the enumeration and detection of bacteriophages in food, water, and beverages. In this chapter, we used the Phi6 SARS-CoV-2 surrogate as an example.