Osong Public Health and Research Perspectives

Original Articles

Specification of Bacteriophage Isolated Against Clinical Methicillin-Resistant Staphylococcus Aureus: Ahmad Nasser, Reza Azizian, Mohsen Tabasi, Jamil Kheirvari Khezerloo, Fatemah Sadeghpour Heravi, Morovat Taheri Kalani, Norkhoda Sadeghifard, Razieh Amini, Iraj Pakzad, Amin Radmanesh, Farid Azizi Jalilian; Osong Public Health Res Perspect. 2019;10(1):20-24. Published online February 28, 2019; DOI: https://doi.org/10.24171/j.phrp.2019.10.1.05

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Objectives

The emergence of resistant bacteria is being increasingly reported around the world, potentially threatening millions of lives. Amongst resistant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) is the most challenging to treat. This is due to emergent MRSA strains and less effective traditional antibiotic therapies to Staphylococcal infections. The use of bacteriophages (phages) against MRSA is a new, potential alternate therapy. In this study, morphology, genetic and protein structure of lytic phages against MRSA have been analysed.

Methods

Isolation of livestock and sewage bacteriophages were performed using 0.4 μm membrane filters. Plaque assays were used to determine phage quantification by double layer agar method. Pure plaques were then amplified for further characterization. Sulfate-polyacrylamide gel electrophoresis and random amplification of polymorphic DNA were run for protein evaluation, and genotyping respectively. Transmission electron microscope was also used to detect the structure and taxonomic classification of phage visually.

Results

Head and tail morphology of bacteriophages against MRSA were identified by transmission electron microscopy and assigned to the Siphoviridae family and the Caudovirales order.

Conclusion

Bacteriophages are the most abundant microorganism on Earth and coexist with the bacterial population. They can destroy bacterial cells successfully and effectively. They cannot enter mammalian cells which saves the eukaryotic cells from lytic phage activity. In conclusion, phage therapy may have many potential applications in microbiology and human medicine with no side effect on eukaryotic cells.

Citations

Citations to this article as recorded by

Characterization of a Vibrio parahaemolyticus-targeting lytic bacteriophage SSJ01 and its application in artificial seawater
Jungu Kang, Yoonjee Chang
Food Science and Biotechnology.2024; 33(6): 1505. CrossRef
Isolation and encapsulation of bacteriophage with chitosan nanoparticles for biocontrol of multidrug-resistant methicillin-resistant Staphylococcus aureus isolated from broiler poultry farms
Mona M. Elsayed, Rasha M. Elkenany, Ayman Y. EL-Khateeb, Nehal M. Nabil, Maram M. Tawakol, Heba M. Hassan
Scientific Reports.2024;[Epub] CrossRef
P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri
Yang W. Huan, Vincenzo Torraca, Russell Brown, Jidapha Fa-arun, Sydney L. Miles, Diego A. Oyarzún, Serge Mostowy, Baojun Wang
ACS Synthetic Biology.2023; 12(3): 709. CrossRef
Staphylococcus aureus Dormancy: Waiting for Insurgency
Ahmad Nasser, Shiva Jahanbakhshi, Mohammad Mehdi Soltan Dallal, Maryam Banar, Azin Sattari-Maraji, Taher Azimi
Current Pharmaceutical Biotechnology.2023; 24(15): 1898. CrossRef
Recent advance on nanoparticles or nanomaterials with anti-multidrug resistant bacteria and anti-bacterial biofilm properties: A systematic review
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Heliyon.2023; 9(11): e22105. CrossRef
Isolation and characterization of lytic bacteriophages from sewage at an egyptian tertiary care hospital against methicillin-resistant Staphylococcus aureus clinical isolates
Safia Samir, Amira El-Far, Hend Okasha, Rania Mahdy, Fatima Samir, Sami Nasr
Saudi Journal of Biological Sciences.2022; 29(5): 3097. CrossRef
Staphylococcus aureus: Biofilm Formation and Strategies Against it
Ahmad Nasser , Mohammad Mehdi Soltan Dallal, Shiva Jahanbakhshi, Taher Azimi, Leila Nikouei
Current Pharmaceutical Biotechnology.2022; 23(5): 664. CrossRef
An Anti-MRSA Phage From Raw Fish Rinse: Stability Evaluation and Production Optimization
Israa M. Abd-Allah, Ghadir S. El-Housseiny, Mohammad Y. Alshahrani, Samar S. El-Masry, Khaled M. Aboshanab, Nadia A. Hassouna
Frontiers in Cellular and Infection Microbiology.2022;[Epub] CrossRef
Molecular mechanisms of Shigella effector proteins: a common pathogen among diarrheic pediatric population
Ahmad Nasser, Mehrdad Mosadegh, Taher Azimi, Aref Shariati
Molecular and Cellular Pediatrics.2022;[Epub] CrossRef
A Metal-Containing NP Approach to Treat Methicillin-Resistant Staphylococcus aureus (MRSA): Prospects and Challenges
Wendy Wai Yeng Yeo, Sathiya Maran, Amanda Shen-Yee Kong, Wan-Hee Cheng, Swee-Hua Erin Lim, Jiun-Yan Loh, Kok-Song Lai
Materials.2022; 15(17): 5802. CrossRef
Electrochemical Biosensors for Pathogen Detection: An Updated Review
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Biosensors.2022; 12(11): 927. CrossRef
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Sara Khosravi, Razieh Amini, Mohammad Reza Arabestani, Seyed Saman Talebi, Farid Azizi Jalilian
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Food and Environmental Virology.2021; 13(4): 433. CrossRef
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Ahmad Nasser, Taher Azimi, Soheila Ostadmohammadi, Samaneh Ostadmohammadi
Microbial Pathogenesis.2020; 148: 104431. CrossRef
Characterization of the Three New Kayviruses and Their Lytic Activity Against Multidrug-Resistant Staphylococcus aureus
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Microorganisms.2019; 7(10): 471. CrossRef

Expression of Recombinant pET22b-LysK-Cysteine/Histidine-Dependent Amidohydrolase/Peptidase Bacteriophage Therapeutic Protein in Escherichia coli BL21 (DE3): Hamed Haddad Kashani, Rezvan Moniri; Osong Public Health Res Perspect. 2015;6(4):256-260. Published online August 31, 2015; DOI: https://doi.org/10.1016/j.phrp.2015.08.001

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Abstract PDF

Objectives
Bacteriophage-encoded endolysins are a group of enzymes that act by digesting the peptidoglycan of bacterial cell walls. LysK has been reported to lyse live staphylococcal cultures. LysK proteins containing only the cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) domain has the capability to show lytic activity against live clinical staphylococcal isolates, including methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to clone and express LysK-CHAP domain in Escherichia coli BL21 (DE3) using pET22b as a secretion vector. The pET22b plasmid was used, which encoded a pelB secretion signal under the control of the strong bacteriophage T7 promoter.
Methods
The E. coli cloning strains DH5α and BL21 (DE3) were grown at 37°C with aeration in the Luria-Bertani medium. A plasmid encoding LysK-CHAP in a pET22b backbone was constructed. The pET22b vector containing LysK-CHAP sequences were digested with NcoI and HindIII restriction enzymes. Cloning accuracy was confirmed by electrophoresis. The pET22b-LysK plasmid was used to transform the E. coli strain BL21. Isopropyl β-d-1-thiogalactopyranoside (IPTG) was added to a final concentration of 1mM to induce T7 RNA polymerase-based expression. Finally, western blot confirmed the expression of target protein.
Results
In this study, after double digestion of pEX and pET22b vectors with HindIII and NcoI, LysK gene was cloned into two HindIII and NcoI sites in pET22b vector, and then transformed to E. coli DH5α. Cloning was confirmed with double digestion and analyzed with agarose gel. The recombinant pET22b-LysK plasmid was transformed to E. coli BL21 and the expression was induced by IPTG. The expression was confirmed by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting method. Observation of a 28.5 kDa band confirmed LysK protein expression.
Conclusion
In the present study, LysK-CHAP domain was successfully cloned and expressed at the pET22b vector and E. coli BL21 (DE3).

Citations

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