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Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction
Non-protein coding RNA (npcRNA) is a functional RNA molecule that is not translated into a protein. Bacterial npcRNAs are structurally diversified molecules, typically 50–200 nucleotides in length. They play a crucial physiological role in cellular networking, including stress responses, replication...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4930178/ https://www.ncbi.nlm.nih.gov/pubmed/27367909 http://dx.doi.org/10.1371/journal.pone.0158736 |
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author | Soo Yean, Cheryl Yeap Selva Raju, Kishanraj Xavier, Rathinam Subramaniam, Sreeramanan Gopinath, Subash C. B. Chinni, Suresh V. |
author_facet | Soo Yean, Cheryl Yeap Selva Raju, Kishanraj Xavier, Rathinam Subramaniam, Sreeramanan Gopinath, Subash C. B. Chinni, Suresh V. |
author_sort | Soo Yean, Cheryl Yeap |
collection | PubMed |
description | Non-protein coding RNA (npcRNA) is a functional RNA molecule that is not translated into a protein. Bacterial npcRNAs are structurally diversified molecules, typically 50–200 nucleotides in length. They play a crucial physiological role in cellular networking, including stress responses, replication and bacterial virulence. In this study, by using an identified npcRNA gene (Sau-02) in Methicillin-resistant Staphylococcus aureus (MRSA), we identified the Gram-positive bacteria S. aureus. A Sau-02-mediated monoplex Polymerase Chain Reaction (PCR) assay was designed that displayed high sensitivity and specificity. Fourteen different bacteria and 18 S. aureus strains were tested, and the results showed that the Sau-02 gene is specific to S. aureus. The detection limit was tested against genomic DNA from MRSA and was found to be ~10 genome copies. Further, the detection was extended to whole-cell MRSA detection, and we reached the detection limit with two bacteria. The monoplex PCR assay demonstrated in this study is a novel detection method that can replicate other npcRNA-mediated detection assays. |
format | Online Article Text |
id | pubmed-4930178 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-49301782016-07-18 Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction Soo Yean, Cheryl Yeap Selva Raju, Kishanraj Xavier, Rathinam Subramaniam, Sreeramanan Gopinath, Subash C. B. Chinni, Suresh V. PLoS One Research Article Non-protein coding RNA (npcRNA) is a functional RNA molecule that is not translated into a protein. Bacterial npcRNAs are structurally diversified molecules, typically 50–200 nucleotides in length. They play a crucial physiological role in cellular networking, including stress responses, replication and bacterial virulence. In this study, by using an identified npcRNA gene (Sau-02) in Methicillin-resistant Staphylococcus aureus (MRSA), we identified the Gram-positive bacteria S. aureus. A Sau-02-mediated monoplex Polymerase Chain Reaction (PCR) assay was designed that displayed high sensitivity and specificity. Fourteen different bacteria and 18 S. aureus strains were tested, and the results showed that the Sau-02 gene is specific to S. aureus. The detection limit was tested against genomic DNA from MRSA and was found to be ~10 genome copies. Further, the detection was extended to whole-cell MRSA detection, and we reached the detection limit with two bacteria. The monoplex PCR assay demonstrated in this study is a novel detection method that can replicate other npcRNA-mediated detection assays. Public Library of Science 2016-07-01 /pmc/articles/PMC4930178/ /pubmed/27367909 http://dx.doi.org/10.1371/journal.pone.0158736 Text en © 2016 Soo Yean et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Soo Yean, Cheryl Yeap Selva Raju, Kishanraj Xavier, Rathinam Subramaniam, Sreeramanan Gopinath, Subash C. B. Chinni, Suresh V. Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title | Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title_full | Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title_fullStr | Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title_full_unstemmed | Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title_short | Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction |
title_sort | molecular detection of methicillin-resistant staphylococcus aureus by non-protein coding rna-mediated monoplex polymerase chain reaction |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4930178/ https://www.ncbi.nlm.nih.gov/pubmed/27367909 http://dx.doi.org/10.1371/journal.pone.0158736 |
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