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PLoS ONE. Isolation and Characterization of φkm18p, a Novel Lytic Phage with Therapeutic Potential against Extensively Drug Resistant Acinetobacter baumannii
[Source: PLoS ONE, full text: (LINK). Abstract, edited.]
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Isolation and Characterization of φkm18p, a Novel Lytic Phage with Therapeutic Potential against Extensively Drug Resistant Acinetobacter baumannii
Gwan-Han Shen<SUP>1</SUP><SUP>,</SUP><SUP>2</SUP><SUP>#</SUP>, Jiun-Ling Wang<SUP>3</SUP><SUP>,</SUP><SUP>4</SUP><SUP>#</SUP>, Fu-Shyan Wen<SUP>5</SUP>, Kai-Ming Chang<SUP>5</SUP>, Chih-Feng Kuo<SUP>6</SUP>, Chun-**** Lin<SUP>7</SUP>, Huei-Ru Luo<SUP>7</SUP>, Chih-Hsin ****<SUP>7</SUP><SUP>*</SUP>
<SUP></SUP>
1 Division of Respiratory and Critical Care Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, Taiwan, ROC, 2 Department of Respiratory Therapy, College of Health Care, China Medical University, Taichung, Taiwan, ROC, 3 School of Chinese Medicine for Post Baccalaureate, I-Shou University, Kaohsiung County, Taiwan, ROC, 4 Department of Internal Medicine, E-Da Hospital, Kaohsiung County, Taiwan, 5 Department of Life Science, National Chung Hsing University, Taichung, Taiwan, ROC, 6 Department of Nursing, I-Shou University, Kaoshiung, Taiwan, ROC, 7 Department of Chemical Engineering, and Institute of Biotechnology and Chemical Engineering, I-Shou University, Kaoshiung, Taiwan, ROC
Abstract
Aims
To isolate phages against extensively drug resistant Acinetobacter baumannii (XDRAB) and characterize the highest lytic capability phage as a model to evaluate the potential on phage therapy.
Methods and Results
Eight phages were isolated from hospital sewage and showed narrow host spectrum. Phage φkm18p was able to effectively lyse the most XDRAB. It has a dsDNA genome of 45 kb in size and hexagonal head of about 59 nm in diameter and no tail. Bacterial population decreased quickly from 10<SUP>8</SUP> CFU ml<SUP>−1</SUP> to 10<SUP>3</SUP> CFU ml<SUP>−1</SUP> in 30 min by φkm18p. The 185 kDa lysis protein encoded by φkm18p genome was detected when the extracted protein did not boil before SDS-PAGE; it showed that the lysis protein is a complex rather than a monomer. Phage φkm18p improved human lung epithelial cells survival rates when they were incubated with A. baumannii. Combination of phages (φkm18p, φTZ1 and φ314) as a cocktail could lyse all genotype-varying XDRAB isolates.
Conclusion
Infections with XDRAB are extremely difficult to treat and development of a phage cocktails therapy could be a therapeutic alternative in the future. Phage φkm18p is a good candidate for inclusion in phage cocktails.
Citation: Shen G-H, Wang J-L, Wen F-S, Chang K-M, Kuo C-F, et al. (2012) Isolation and Characterization of φkm18p, a Novel Lytic Phage with Therapeutic Potential against Extensively Drug Resistant Acinetobacter baumannii. PLoS ONE 7(10): e46537. doi:10.1371/journal.pone.0046537
Editor: Brad Spellberg, Los Angeles Biomedical Research Institute, United States of America
Received: April 30, 2012; Accepted: August 31, 2012; Published: October 5, 2012
Copyright: ? 2012 Shen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by grants from the National Science Council, Republic of China (No. NSC 94-2311-B-214-001) and I-Shou University (ISU93-02-17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
* E-mail: chhung@isu.edu.tw
# These authors contributed equally to this work.
-Gwan-Han Shen<SUP>1</SUP><SUP>,</SUP><SUP>2</SUP><SUP>#</SUP>, Jiun-Ling Wang<SUP>3</SUP><SUP>,</SUP><SUP>4</SUP><SUP>#</SUP>, Fu-Shyan Wen<SUP>5</SUP>, Kai-Ming Chang<SUP>5</SUP>, Chih-Feng Kuo<SUP>6</SUP>, Chun-**** Lin<SUP>7</SUP>, Huei-Ru Luo<SUP>7</SUP>, Chih-Hsin ****<SUP>7</SUP><SUP>*</SUP>
<SUP></SUP>
1 Division of Respiratory and Critical Care Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, Taiwan, ROC, 2 Department of Respiratory Therapy, College of Health Care, China Medical University, Taichung, Taiwan, ROC, 3 School of Chinese Medicine for Post Baccalaureate, I-Shou University, Kaohsiung County, Taiwan, ROC, 4 Department of Internal Medicine, E-Da Hospital, Kaohsiung County, Taiwan, 5 Department of Life Science, National Chung Hsing University, Taichung, Taiwan, ROC, 6 Department of Nursing, I-Shou University, Kaoshiung, Taiwan, ROC, 7 Department of Chemical Engineering, and Institute of Biotechnology and Chemical Engineering, I-Shou University, Kaoshiung, Taiwan, ROC
Abstract
Aims
To isolate phages against extensively drug resistant Acinetobacter baumannii (XDRAB) and characterize the highest lytic capability phage as a model to evaluate the potential on phage therapy.
Methods and Results
Eight phages were isolated from hospital sewage and showed narrow host spectrum. Phage φkm18p was able to effectively lyse the most XDRAB. It has a dsDNA genome of 45 kb in size and hexagonal head of about 59 nm in diameter and no tail. Bacterial population decreased quickly from 10<SUP>8</SUP> CFU ml<SUP>−1</SUP> to 10<SUP>3</SUP> CFU ml<SUP>−1</SUP> in 30 min by φkm18p. The 185 kDa lysis protein encoded by φkm18p genome was detected when the extracted protein did not boil before SDS-PAGE; it showed that the lysis protein is a complex rather than a monomer. Phage φkm18p improved human lung epithelial cells survival rates when they were incubated with A. baumannii. Combination of phages (φkm18p, φTZ1 and φ314) as a cocktail could lyse all genotype-varying XDRAB isolates.
Conclusion
Infections with XDRAB are extremely difficult to treat and development of a phage cocktails therapy could be a therapeutic alternative in the future. Phage φkm18p is a good candidate for inclusion in phage cocktails.
Citation: Shen G-H, Wang J-L, Wen F-S, Chang K-M, Kuo C-F, et al. (2012) Isolation and Characterization of φkm18p, a Novel Lytic Phage with Therapeutic Potential against Extensively Drug Resistant Acinetobacter baumannii. PLoS ONE 7(10): e46537. doi:10.1371/journal.pone.0046537
Editor: Brad Spellberg, Los Angeles Biomedical Research Institute, United States of America
Received: April 30, 2012; Accepted: August 31, 2012; Published: October 5, 2012
Copyright: ? 2012 Shen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by grants from the National Science Council, Republic of China (No. NSC 94-2311-B-214-001) and I-Shou University (ISU93-02-17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
* E-mail: chhung@isu.edu.tw
# These authors contributed equally to this work.
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