title: Molecular Detection of Methicillin-Resistant Staphylococcus aureus by Non-Protein Coding RNA-Mediated Monoplex Polymerase Chain Reaction.
abstract: 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.
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong, Malaysia.
authors.forename: Cheryl Yeap
authors.initials: CY
authors.lastname: Soo Yean
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong, Malaysia.
authors.forename: Kishanraj
authors.initials: K
authors.lastname: Selva Raju
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong, Malaysia.
authors.forename: Rathinam
authors.initials: R
authors.lastname: Xavier
authors.affiliation: School of Biological Sciences, Universiti Sains Malaysia, Penang, Malaysia.
authors.forename: Sreeramanan
authors.initials: S
authors.lastname: Subramaniam
authors.affiliation: Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, Kangar, Perlis, Malaysia.|School of Bioprocess Engineering, Universiti Malaysia Perlis, Arau, Perlis, Malaysia.
authors.forename: Subash C B
authors.initials: SC
authors.lastname: Gopinath
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Bedong, Malaysia.
authors.forename: Suresh V
authors.initials: SV
authors.lastname: Chinni
mesh_terms: D023281:Genomics; D055624:Methicillin-Resistant Staphylococcus aureus / Q000166:cytology / Q000235:genetics* / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D022661:RNA, Untranslated / Q000235:genetics*
chemical_list: D022661:RNA, Untranslated