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A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections
Background and Objective: Wound infections are often difficult to treat due to various bacterial pathogens. Pseudomonas aeruginosa is one of the common invaders of open wounds. Precise diagnosis of this etiological agent in wound infections is of critical importance particularly in treatment of prob...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Professional Medical Publicaitons
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3817763/ https://www.ncbi.nlm.nih.gov/pubmed/24353667 |
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author | Salman, Muhammad Ali, Aamir Haque, Abdul |
author_facet | Salman, Muhammad Ali, Aamir Haque, Abdul |
author_sort | Salman, Muhammad |
collection | PubMed |
description | Background and Objective: Wound infections are often difficult to treat due to various bacterial pathogens. Pseudomonas aeruginosa is one of the common invaders of open wounds. Precise diagnosis of this etiological agent in wound infections is of critical importance particularly in treatment of problematic cases. The existing diagnostic methods have certain limitations particularly related to specificity. Our objective was to to establish a comprehensive and reliable multiplex PCR to confirm diagnosis of P. aeruginosa. Methods: A multiplex PCR test was developed for rapid and comprehensive identification of P. aeruginosa. Four highly specific genes were targeted simultaneously for detection of genus, species and exotoxin production (16S rDNA, gyrB, oprL and ETA) in P. aeruginosa; additionally one internal control gene (invA) of Salmonella was used. The specificity of the multiplex PCR was confirmed using internal and negative controls. Amplified fragments were confirmed by restriction analysis and DNA sequencing. Results: The developed method was applied on 40 morphologically suspected P. aeruginosa isolates (from 200 pus samples) and 18 isolates were confirmed as P. aeruginosa. In comparison, only 12 could be identified biochemically. Conclusions: Combination of the four reported genes in multiplex PCR provided more confident and comprehensive detection of P. aeruginosa which is applicable for screening of wound infections and assisting treatment strategy. |
format | Online Article Text |
id | pubmed-3817763 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Professional Medical Publicaitons |
record_format | MEDLINE/PubMed |
spelling | pubmed-38177632013-12-18 A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections Salman, Muhammad Ali, Aamir Haque, Abdul Pak J Med Sci Original Article Background and Objective: Wound infections are often difficult to treat due to various bacterial pathogens. Pseudomonas aeruginosa is one of the common invaders of open wounds. Precise diagnosis of this etiological agent in wound infections is of critical importance particularly in treatment of problematic cases. The existing diagnostic methods have certain limitations particularly related to specificity. Our objective was to to establish a comprehensive and reliable multiplex PCR to confirm diagnosis of P. aeruginosa. Methods: A multiplex PCR test was developed for rapid and comprehensive identification of P. aeruginosa. Four highly specific genes were targeted simultaneously for detection of genus, species and exotoxin production (16S rDNA, gyrB, oprL and ETA) in P. aeruginosa; additionally one internal control gene (invA) of Salmonella was used. The specificity of the multiplex PCR was confirmed using internal and negative controls. Amplified fragments were confirmed by restriction analysis and DNA sequencing. Results: The developed method was applied on 40 morphologically suspected P. aeruginosa isolates (from 200 pus samples) and 18 isolates were confirmed as P. aeruginosa. In comparison, only 12 could be identified biochemically. Conclusions: Combination of the four reported genes in multiplex PCR provided more confident and comprehensive detection of P. aeruginosa which is applicable for screening of wound infections and assisting treatment strategy. Professional Medical Publicaitons 2013 /pmc/articles/PMC3817763/ /pubmed/24353667 Text en This is an Open Access article distributed under the terms of the Creative Commons Attribution License, (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Salman, Muhammad Ali, Aamir Haque, Abdul A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title | A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title_full | A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title_fullStr | A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title_full_unstemmed | A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title_short | A novel multiplex PCR for detection of Pseudomonas aeruginosa: A major cause of wound infections |
title_sort | novel multiplex pcr for detection of pseudomonas aeruginosa: a major cause of wound infections |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3817763/ https://www.ncbi.nlm.nih.gov/pubmed/24353667 |
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