AI Article Synopsis

  • The study focuses on improving the quantification of phages for personalized therapy and industrial use, highlighting that the traditional culture-based method is accurate but time-consuming.
  • A DNase treatment before using quantitative polymerase chain reaction (qPCR) is proposed to enhance the accuracy of phage quantification, though qPCR still tends to overestimate compared to the gold standard method.
  • Findings indicate that the differences in phage quantification methods vary based on the age of the phage stocks, and electron microscopy confirmed the presence of free DNA influencing the results.

Article Abstract

To use phages in a personalized therapy and industrial applications, an accurate quantification is needed. The gold standard method, namely the culture-based double agar overlay (DAO) method, provides an accurate estimate of the number of infectious phages but is laborious and time-intensive. Quantitative polymerase chain reaction (qPCR) can be used as a fast alternative but tends to overestimate the number of infectious phage particles. Here we describe the use of a DNase treatment before quantification of the Staphylococcus aureus phage ISP with qPCR to obtain a more accurate estimate of the number of infectious phage particles. We showed that DNase treatment results in a significant decrease of the concentration when measured with qPCR although for two out of three tested ISP phage stocks, there was still a significant difference with the DAO method. We also showed that the discrepancy between quantification with qPCR and the DAO method is dependent on the storage period of the phage stock, with a larger discrepancy for older stocks. Additionally, we used negative contrast immune electron microscopy to confirm the presence of DNA in the medium of the phage stock and the impact of the DNase treatment on the free DNA.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11614264PMC
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0313774PLOS

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