scholarly journals Phages for Phage Therapy: Isolation, Characterization, and Host Range Breadth

2019 ◽  
Vol 12 (1) ◽  
pp. 35 ◽  
Author(s):  
Paul Hyman
Keyword(s):  
Host Range ◽  
Phage Therapy ◽  
Bacterial Pathogen ◽  
Broad Host Range ◽  
Toxin Genes

For a bacteriophage to be useful for phage therapy it must be both isolated from the environment and shown to have certain characteristics beyond just killing strains of the target bacterial pathogen. These include desirable characteristics such as a relatively broad host range and a lack of other characteristics such as carrying toxin genes and the ability to form a lysogen. While phages are commonly isolated first and subsequently characterized, it is possible to alter isolation procedures to bias the isolation toward phages with desirable characteristics. Some of these variations are regularly used by some groups while others have only been shown in a few publications. In this review I will describe (1) isolation procedures and variations that are designed to isolate phages with broader host ranges, (2) characterization procedures used to show that a phage may have utility in phage therapy, including some of the limits of such characterization, and (3) results of a survey and discussion with phage researchers in industry and academia on the practice of characterization of phages.

scholarly journals 1615. Isolation of Lytic Bacteriophages with Broad Host Range Activity Against Pseudomonas aeruginosa Strains Isolated from Respiratory Samples from Cystic Fibrosis Patients Intended for Therapeutic Application

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S801-S801
Author(s):  
Jose Alexander ◽  
Daniel Navas ◽  
Marly Flowers ◽  
Angela Charles ◽  
Amy Carr
Keyword(s):  
Cystic Fibrosis ◽  
Host Range ◽  
Phage Therapy ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Plaque Morphology ◽  
Broad Host Range ◽  
Chronic Infections ◽  
Clinically Significant ◽  
Host Target

Abstract Background With the rise of the antimicrobial resistance between different genera and species of bacteria, Phage Therapy is becoming a more realistic and accessible option for patients with limited or no antimicrobial options. Being able to have rapid access to a collection of clinical active phages is key for rapid implementation of phage therapy. The Microbiology Department at AdventHealth Orlando is performing routine screening of environmental and patient samples for isolation of phages against non-fermenting Gram negative bacteria to develop a Phage Bank. Methods Protocols for phage isolation from environmental sources such as lakes, rivers and sewers and clinical samples were developed. A series of respiratory, throat, stool and urine samples were processed following an internal protocol that includes centrifugation, filtration and enrichment. Clinical samples were centrifugated for 10 minutes, filtered using 0.45µm centrifugation filters, seeded with targeted host bacteria (clinical isolates) and incubated at 35°C for 24 hours. The enriched samples were centrifugated and filtered for a final phage enriched solution. Screening and isolation were performed using the Gracia method over trypticase soybean agar (TSA) for plaque morphology and quantification. Host range screening of other clinical isolates of P. aeruginosa was performed using the new isolated and purified phages. Results 4 lytic phages against clinical strains of P. aeruginosa from patient with diagnosis of cystic fibrosis (CF), were isolated and purified from 4 different respiratory samples, including sputum and bronchial alveolar lavage. All phages showed phenotypical characteristics of lytic activity. 1 phage was active against 4 strains of P. aeruginosa, 1 phage was active against 2 strains of P. aeruginosa and the remaining 2 phages were active only against the initial host target strain. Conclusion With this study we demonstrated the potential use of clinical samples as source for isolating active bacteriophages against clinically significant bacteria strains. Clinical samples from vulnerable population of patients with chronic infections are part of our routine “phage-hunting” process to stock and grow our Phage Bank project for future clinical use. Disclosures All Authors: No reported disclosures

scholarly journals Development of the Automated Primer Design Workflow Uniqprimer and Diagnostic Primers for the Broad-Host-Range Plant Pathogen Dickeya dianthicola

Plant Disease ◽  
2019 ◽  
Vol 103 (11) ◽  
pp. 2893-2902 ◽  
Author(s):  
Shaista Karim ◽  
R. Ryan McNally ◽  
Afnan S. Nasaruddin ◽  
Alexis DeReeper ◽  
Ramil P. Mauleon ◽  
...  
Keyword(s):  
Host Range ◽  
Bacterial Pathogen ◽  
The United States ◽  
Broad Host Range ◽  
Software Pipeline ◽  
Widespread Occurrence ◽  
Field Samples ◽  
Primer Sets ◽  
Genome Analyses ◽  
User Friendly

Uniqprimer, a software pipeline developed in Python, was deployed as a user-friendly internet tool in Rice Galaxy for comparative genome analyses to design primer sets for PCRassays capable of detecting target bacterial taxa. The pipeline was trialed with Dickeya dianthicola, a destructive broad-host-range bacterial pathogen found in most potato-growing regions. Dickeya is a highly variable genus, and some primers available to detect this genus and species exhibit common diagnostic failures. Upon uploading a selection of target and nontarget genomes, six primer sets were rapidly identified with Uniqprimer, of which two were specific and sensitive when tested with D. dianthicola. The remaining four amplified a minority of the nontarget strains tested. The two promising candidate primer sets were trialed with DNA isolated from 116 field samples from across the United States that were previously submitted for testing. D. dianthicola was detected in 41 samples, demonstrating the applicability of our detection primers and suggesting widespread occurrence of D. dianthicola in North America.

scholarly journals Genetic Characterization of Broad-Host-Range IncQ Plasmids Harboring blaVEB-18 in Vibrio Species

2017 ◽  
Vol 61 (7) ◽  
Author(s):  
Ruichao Li ◽  
Lianwei Ye ◽  
Zhiwei Zheng ◽  
Edward Wai Chi Chan ◽  
Sheng Chen
Keyword(s):  
Host Range ◽  
Genetic Characterization ◽  
Vibrio Species ◽  
Broad Host Range ◽  
Incq Plasmids

scholarly journals Genomic and Proteomic Characterization of the Broad-Host-Range Salmonella Phage PVP-SE1: Creation of a New Phage Genus

2011 ◽  
Vol 85 (21) ◽  
pp. 11265-11273 ◽  
Author(s):  
S. B. Santos ◽  
A. M. Kropinski ◽  
P.-J. Ceyssens ◽  
H.- W. Ackermann ◽  
A. Villegas ◽  
...  
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Salmonella Phage

Characterization of Gram-positive broad host-range plasmids carrying a thermophilic replicon

1991 ◽  
Vol 142 (4) ◽  
pp. 389-396 ◽  
Author(s):  
E De Rossi ◽  
P Brigidi ◽  
M Rossi ◽  
D Matteuzzi ◽  
G Riccardi
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Gram Positive ◽  
Broad Host Range Plasmids

scholarly journals JD419, a Staphylococcus aureus Phage With a Unique Morphology and Broad Host Range

2021 ◽  
Vol 12 ◽  
Author(s):  
Tingting Feng ◽  
Sebastian Leptihn ◽  
Ke Dong ◽  
Belinda Loh ◽  
Yan Zhang ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Host Range ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Complete Characterization ◽  
Resistant Bacteria ◽  
Broad Host Range ◽  
Gene Engineering ◽  
Mrsa Strains

Phage therapy represents a possible treatment option to cure infections caused by multidrug-resistant bacteria, including methicillin and vancomycin-resistant Staphylococcus aureus, to which most antibiotics have become ineffective. In the present study, we report the isolation and complete characterization of a novel phage named JD219 exhibiting a broad host range able to infect 61 of 138 clinical strains of S. aureus tested, which included MRSA strains as well. The phage JD419 exhibits a unique morphology with an elongated capsid and a flexible tail. To evaluate the potential of JD419 to be used as a therapeutic phage, we tested the ability of the phage particles to remain infectious after treatment exceeding physiological pH or temperature. The activity was retained at pH values of 6.0–8.0 and below 50°C. As phages can contain virulence genes, JD419’s complete genome was sequenced. The 45509 bp genome is predicted to contain 65 ORFs, none of which show homology to any known virulence or antibiotic resistance genes. Genome analysis indicates that JD419 is a temperate phage, despite observing rapid replication and lysis of host strains. Following the recent advances in synthetic biology, JD419 can be modified by gene engineering to remove prophage-related genes, preventing potential lysogeny, in order to be deployed as a therapeutic phage.

scholarly journals Isolation and characterization of novel broad host range bacteriophages of Vibrio cholerae O1 from Bengal

2018 ◽  
Vol 10 (2) ◽  
pp. 84
Author(s):  
BanwarilalL Sarkar ◽  
Sounak Sarkar ◽  
Mayukh Das ◽  
TusharSuvra Bhowmick ◽  
Hemanta Koley ◽  
...  
Keyword(s):  
Host Range ◽  
Vibrio Cholerae ◽  
Broad Host Range ◽  
Isolation And Characterization ◽  
Vibrio Cholerae O1
Sign in / Sign up

Export Citation Format

Share Document