Use of the polymerase chain reaction and 16S rRNA sequences for the rapid detection of Brochothrix spp. in foods
- 11 March 1993
- journal article
- Published by Wiley in Journal of Applied Bacteriology
- Vol. 74 (3) , 260-267
- https://doi.org/10.1111/j.1365-2672.1993.tb03024.x
Abstract
Oligonucleotide primers were designed against rRNA sequences to give a DNA-based PCR assay for the rapid identification/detection of Brochothrix spp. The PCR products could be confirmed by hybridization to an internal oligonucleotide probe. The method successfully and sensitively detected/identified these organisms in pure cultures but was of limited value as a detection method because the detection sensitivity, in relation to conventional plate counts, varied and the assay sensitivity was reduced in the presence of staphylococci. Furthermore, sensitivity was also lost when the assay was applied directly to meat samples. However, a separation step using a lectin (from Agaricus bisporus) immobilized on magnetic beads prior to the PCR assay, allowed the direct detection of low numbers (> 10(2) cfu g-1) of Brochothrix in meat samples within a working day.Keywords
This publication has 29 references indexed in Scilit:
- The interrelationships of all major groups of Platyhelminthes: phylogenetic evidence from morphology and moleculesBiological Journal of the Linnean Society, 1999
- The use of immobilized lectins in the separation of Staphylococcus aureus, Escherichia coli, Listeria and Salmonella spp. from pure cultures and foodsJournal of Applied Bacteriology, 1992
- Use of the polymerase chain reaction and oligonucleotide probes for the rapid detection and identification of Carnobacterium species from meatJournal of Applied Bacteriology, 1992
- Detection and identification of Listeria monocytogenes in cooked sausage products and in milk by in vitro amplification of haemolysin gene fragmentsJournal of Applied Bacteriology, 1991
- Phylogenetic Analysis of the Genus Listeria Based on Reverse Transcriptase Sequencing of 16S rRNAInternational Journal of Systematic and Evolutionary Microbiology, 1991
- Use of the polymerase chain reaction for direct detection of Listeria monocytogenes in soft cheeseJournal of Applied Bacteriology, 1991
- Multiplex PCR amplification and immobilized capture probes for detection of bacterial pathogens and indicators in waterMolecular and Cellular Probes, 1990
- DNA-rRNA Hybridization Studies among Brochothrix spp. and Some Other Gram-Positive BacteriaInternational Journal of Systematic and Evolutionary Microbiology, 1990
- Detection and identification of mycobacteria by amplification of rRNAJournal of Clinical Microbiology, 1990
- Rapid extraction of bacterial genomic DNA with guanidium thiocyanateLetters in Applied Microbiology, 1989