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Application of a molecular beacon based real-time isothermal amplification (MBRTIA) technology for simultaneous detection of Bacillus cereus and Staphylococcus aureus

Author:
Mandappa, I. M., Joglekar, Prasanna, Manonmani, H. K.
Source:
Journal of food science and technology 2015 v.52 no.7 pp. 4642-4646
ISSN:
0022-1155
Subject:
Bacillus cereus, DNA, DNA-directed DNA polymerase, Staphylococcus aureus, food pathogens, genes, nucleotide sequences, polymerase chain reaction, temperature, virulence
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 10¹ 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.
Agid:
4406878