A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis

Khairnar, Krishna; Sanmukh, Swapnil Ganesh; Chandekar, Rajshree; Paunikar, Waman

doi:10.1016/j.jviromet.2014.02.028

Cited by 6 publications

(6 citation statements)

References 27 publications

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“…Isolation of phages against all the E. coli strains was carried out in the TSA medium by pour plating as per Lillehaug protocol with some modification [40,41]. For the pour plating assay, 100 µL of bacterial culture at OD 550 ~0.1 (approximately 1 × 10 8 CFU/mL) was inoculated with 10 µL of processed clinical lysate separately and incubated at room temperature for 5-10 min before plating.…”

Section: Processing Of Clinical Samples For Phage Isolationmentioning

confidence: 99%

“…Later, 10% v/v chloroform was added to the culture with further incubation for more than 30 min at 37 • C, and then the culture was kept at 4 • C without shaking overnight. Finally, the bacterial debris was removed by centrifugation at 5000× g for 20 min at 4 • C, and the supernatant obtained was filtered through a 0.22 mm Millipore TM filter and stored at 4 • C as a phage stock [41].…”

Section: Phage Propagation and Lysate Preparationmentioning

confidence: 99%

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Accessing the In Vivo Efficiency of Clinically Isolated Phages against Uropathogenic and Invasive Biofilm-Forming Escherichia coli Strains for Phage Therapy

Admella

Moya-Andérico

Fehér

et al. 2023

Cells

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Escherichia coli is one of the most common members of the intestinal microbiota. Many of its strains are associated with various inflammatory infections, including urinary or gut infections, especially when displaying antibiotic resistance or in patients with suppressed immune systems. According to recent reports, the biofilm-forming potential of E. coli is a crucial factor for its increased resistance against antibiotics. To overcome the limitations of using antibiotics against resistant E. coli strains, the world is turning once more towards bacteriophage therapy, which is becoming a promising candidate amongst the current personalized approaches to target different bacterial infections. Although matured and persistent biofilms pose a serious challenge to phage therapy, they can still become an effective alternative to antibiotic treatment. Here, we assess the efficiency of clinically isolated phages in phage therapy against representative clinical uropathogenic and invasive biofilm-forming E. coli strains. Our results demonstrate that irrespective of host specificity, bacteriophages producing clear plaques with a high burst size, and exhibiting depolymerizing activity, are good candidates against biofilm-producing E. coli pathogens as verified from our in vitro and in vivo experiments using Galleria mellonella where survival was significantly increased for phage-therapy-treated larvae.

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Section: Processing Of Clinical Samples For Phage Isolationmentioning

confidence: 99%

Section: Phage Propagation and Lysate Preparationmentioning

confidence: 99%

Accessing the In Vivo Efficiency of Clinically Isolated Phages against Uropathogenic and Invasive Biofilm-Forming Escherichia coli Strains for Phage Therapy

Admella

Moya-Andérico

Fehér

et al. 2023

Cells

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“…The cells that present the greatest damage in the DNA show increased migration of the tapes of the genetic material toward the anode (Singh et al, 1988). Khairnar et al (2014) compared the comet assay with other genotoxic tests in order to detect lysis of bacteriophage-mediated bacterial cells. They observed that the comet assay is a simple, practical and economical procedure, demonstrating efficacy in the analysis, allowing the visualization of lysed cells (Khainar et al, 2014).…”

Section: B) Genotoxicity Assays Comet Assaymentioning

confidence: 99%

Análise Do Percentual De Colágeno Na Dentina Humana Entre Diferentes Etnias, Gêneros E Idades

Furtado¹,

Santos²,

Santos³

et al. 2020

Comunicação Científica E Técnica Em Odontologia 4

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Todo o conteúdo deste livro está licenciado sob uma Licença de Atribuição Creative Commons. Atribuição 4.0 Internacional (CC BY 4.0).O conteúdo dos artigos e seus dados em sua forma, correção e confiabilidade são de responsabilidade exclusiva dos autores. Permitido o download da obra e o compartilhamento desde que sejam atribuídos créditos aos autores, mas sem a possibilidade de alterá-la de nenhuma forma ou utilizá-la para fins comerciais.

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“…[27][28][29] Still, the method leads to bacteriophage detection in bacteriophage-mediated cell lysis. 30 Thus, the employability of the CA is very wide, demonstrating that the methodology is still present in mutagenesis and ecogenotoxicity studies. The variety of studies involving this method allows us considering it as an essential methodology in the areas of biochemistry, genetics, molecular biology, pharmacology, toxicology, and pharmaceutics.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of DNA damage and toxicological methodology development: A bibliometric study

et al. 2020

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Genomic instability is a risk to organism health detected by methods such as the comet assay (CA). It is a highly sensitive and versatile method to detect low levels of DNA damage in a wide range of cells from humans as well as from other species as compared to other methods with the same proposal. CA is a powerful DNA damage analysis tool and its applicability extends to the genotoxicity analysis of, that is, drugs and carcinogenic substances. This study analyzed papers employing CA in the Scopus database in order to assess its scientific importance, employability, and trends by evaluating: number of articles per year, total citations and per year, country of publication and their clusters, clusters of authors, most frequently abstracts terms, name of journal, affiliations, country of publication, subject area, relevant keywords compared to citation clusters, and impact factor (IF) CiteScore. It was retrieved 13,828 articles from 1990 to 2018, with a peak in 2014 and a decline from 2015 to 2018. Four author clusters from China, United States, India, and Brazil were identified, countries presenting the greatest number of publications. China presented the most recent scientific advances in the field. It was also detected nine clusters of themes, and a positive correlation between publications, citations, and the IF. There are full employability and versatility in the use of the method. Currently, there is an advance in Chinese scientific production on the subject, and there is greater use of the method on oxidative damage researches.

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A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis

Cited by 6 publications

References 27 publications

Accessing the In Vivo Efficiency of Clinically Isolated Phages against Uropathogenic and Invasive Biofilm-Forming Escherichia coli Strains for Phage Therapy

Accessing the In Vivo Efficiency of Clinically Isolated Phages against Uropathogenic and Invasive Biofilm-Forming Escherichia coli Strains for Phage Therapy

Análise Do Percentual De Colágeno Na Dentina Humana Entre Diferentes Etnias, Gêneros E Idades

Evaluation of DNA damage and toxicological methodology development: A bibliometric study

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