title: Application of a molecular beacon based real-time isothermal amplification (MBRTIA) technology for simultaneous detection of Bacillus cereus and Staphylococcus aureus.
abstract: A multiplex real-time isothermal amplification assay was developed using molecular beacons for the detection of Bacillus cereus and Staphylococcus aureus by targeting four important virulence genes. A correlation between targeting highly accessible DNA sequences and isothermal amplification based molecular beacon efficiency and sensitivity was demonstrated using phi(Φ)29 DNA polymerase at a constant isothermal temperature of 30 °C. It was very selective and consistently detected down to 10(1) copies of DNA. The specificity and sensitivity of this assay, when tested with pure culture were high, surpassing those of currently used PCR assays for the detection of these organisms. The molecular beacon based real-time isothermal amplification (MBRTIA) assay could be carried out entirely in 96 well plates or well strips, enabling a rapid and high-throughput detection of food borne pathogens.
authors.affiliation: Fermentation Technology and Bioengineering Department, CSIR-Central Food Technological Research Institute, Mysore, 570 020 Karnataka India.
authors.forename: I M
authors.initials: IM
authors.lastname: Mandappa
authors.affiliation: Fermentation Technology and Bioengineering Department, CSIR-Central Food Technological Research Institute, Mysore, 570 020 Karnataka India.
authors.forename: Prasanna
authors.initials: P
authors.lastname: Joglekar
authors.affiliation: Fermentation Technology and Bioengineering Department, CSIR-Central Food Technological Research Institute, Mysore, 570 020 Karnataka India.
authors.forename: H K
authors.initials: HK
authors.lastname: Manonmani
keywords: B. cereus; Detection; Food borne pathogens; Molecular beacon; S. aureus