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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...

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Autores principales: Soo Yean, Cheryl Yeap, Selva Raju, Kishanraj, Xavier, Rathinam, Subramaniam, Sreeramanan, Gopinath, Subash C. B., Chinni, Suresh V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
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.
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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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