A rapid and simple protocol for the isolation of bacteriophages from coastal organisms

Echeverría-Vega, Alex; Morales-Vicencio, Pablo; Saez-Saavedra, Camila; Gordillo-Fuenzalida, Felipe; Araya, Rubén

doi:10.1016/j.mex.2019.11.003

Cited by 20 publications

(17 citation statements)

References 7 publications

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“…Many published protocols describe the isolation of desired phages from the environment, each with advantages and disadvantages. The reported yield of most protocols is in the range of 10 4 to 10 9 PFU/ml compared to the protocol described here, where the yield is as high as 10 14 PFU/ml (Echeverría‐Vega et al., 2019; Hallajzadeh, Mojtahedi, Mahabadi, & Amirmozafari, 2019; Sangha, Kumar, Agrawal, Deka, & Verma, 2014). We have shown that sample pooling has multiple advantages over individual sample processing.…”

Section: Commentarymentioning

confidence: 62%

Simple Two‐step, High Yield Protocol for Isolation and Amplification of Bacteriophages Against Methicillin‐resistant Staphylococcus Aureus (MRSA)

Khan

Joshi

2022

Current Protocols

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Bacteriophages are bacteria‐targeting viruses that may prove useful as therapeutic agents against multidrug‐resistant bacterial strains. Though phage therapy is a century‐old concept, there is very limited progress on its therapeutic application due to the rapid expansion of antibiotics portfolios in the last few decades. However, the emergence of multidrug‐resistant organisms has brought our attention back to bacteriophages. The first step towards developing effective phage therapy against multidrug‐resistant bacteria is isolation, amplification, and purification of specific bacteriophages. There are many reported protocols for isolating host‐specific bacteriophages from the environment. However, most of them are complex, multistep, low‐yielding, resource‐intensive protocols, requiring elaborate laboratory setup. We have demonstrated a simple two‐step, high‐yielding protocol for isolating and amplifying bacteriophages against methicillin‐resistant Staphylococcus aureus (MRSA). We have shown that mixing various environmental samples (i.e., sample pooling) and phage amplification at two different temperatures significantly enhance the yield of MRSA phages. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Preparation of water sample filtrate for isolation of bacteriophages Basic Protocol 2: Bacterial strain and culture conditions Basic Protocol 3: Native bacteriophage count in water sample filtrate Basic Protocol 4: Isolation and enrichment of MRSA‐specific bacteriophages Basic Protocol 5: Quantification of bacteriophages by drop cast method Basic Protocol 6: Effect of incubation temperature and heat shock on bacteriophage yield

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Section: Commentarymentioning

confidence: 62%

Simple Two‐step, High Yield Protocol for Isolation and Amplification of Bacteriophages Against Methicillin‐resistant Staphylococcus Aureus (MRSA)

Khan

Joshi

2022

Current Protocols

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“…On the contrary, a study done by Mattila et al, ( Mattila et al, 2015 ) in Finland showed that it was not easy to isolate phages against MRSA from sewage drains. The efficacious recovery of bacteriophages capable of killing precise pathogenic bacteria is dependent on host density and surrounding environmental conditions ( Echeverría-Vega et al, 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of lytic bacteriophages from sewage at an egyptian tertiary care hospital against methicillin-resistant Staphylococcus aureus clinical isolates

Samir

El-Far

Nasr

et al. 2022

Saudi Journal of Biological Sciences

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“…Echeverría-Vega et al [23] used a straightforward protocol for the isolation of bacteriophages from coastal organisms. They also validated the protocol for the isolation of lytic bacteriophages for the fish pathogen bacterium Vibrio ordalii.…”

Section: Phages As Pharmaceuticals 21 Phage Isolation and Enrichmentmentioning

confidence: 99%

Bacteriophage Therapy of Bacterial Infections: The Rediscovered Frontier

2021

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Antibiotic-resistant infections present a serious health concern worldwide. It is estimated that there are 2.8 million antibiotic-resistant infections and 35,000 deaths in the United States every year. Such microorganisms include Acinetobacter, Enterobacterioceae, Pseudomonas, Staphylococcus and Mycobacterium. Alternative treatment methods are, thus, necessary to treat such infections. Bacteriophages are viruses of bacteria. In a lytic infection, the newly formed phage particles lyse the bacterium and continue to infect other bacteria. In the early 20th century, d’Herelle, Bruynoghe and Maisin used bacterium-specific phages to treat bacterial infections. Bacteriophages are being identified, purified and developed as pharmaceutically acceptable macromolecular “drugs,” undergoing strict quality control. Phages can be applied topically or delivered by inhalation, orally or parenterally. Some of the major drug-resistant infections that are potential targets of pharmaceutically prepared phages are Pseudomonas aeruginosa, Mycobacterium tuberculosis and Acinetobacter baumannii.

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A rapid and simple protocol for the isolation of bacteriophages from coastal organisms

Abstract: Graphical abstract

Cited by 20 publications

References 7 publications

Simple Two‐step, High Yield Protocol for Isolation and Amplification of Bacteriophages Against Methicillin‐resistant Staphylococcus Aureus (MRSA)

Simple Two‐step, High Yield Protocol for Isolation and Amplification of Bacteriophages Against Methicillin‐resistant Staphylococcus Aureus (MRSA)

Isolation and characterization of lytic bacteriophages from sewage at an egyptian tertiary care hospital against methicillin-resistant Staphylococcus aureus clinical isolates

Bacteriophage Therapy of Bacterial Infections: The Rediscovered Frontier

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