Use of the polymerase chain reaction and oligonucleotide probes for the rapid detection and identification of Carnobacterium species from meat

The polymerase chain reaction (PCR) was used selectively to amplify specific rDNA sequences of Carnobacterium divergens, C. mobile, C. piscicola and C. gallinarum in purified DNA extracts, crude cell lysates and food samples. The PCR products were visualized by agarose gel electrophoresis and identi...

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Bibliographic Details
Published inJournal of applied bacteriology Vol. 72; no. 4; pp. 294 - 301
Main Authors Brooks, J.L, Moore, A.S, Patchett, R.A, Collins, M.D, Kroll, R.G
Format Journal Article
LanguageEnglish
Published England 01.04.1992
Subjects
bacteria
Base Sequence
biochemical techniques
DNA
DNA, Bacterial - genetics
DNA, Ribosomal - genetics
Food Microbiology
Gram-Positive Asporogenous Rods - classification
Gram-Positive Asporogenous Rods - genetics
Gram-Positive Asporogenous Rods - isolation & purification
identification
Lactobacillaceae - classification
Lactobacillaceae - genetics
Lactobacillaceae - isolation & purification
meat
Meat - microbiology
Molecular Sequence Data
Nucleic Acid Hybridization
Oligonucleotide Probes
Polymerase Chain Reaction
rapid methods
RNA, Bacterial - genetics
RNA, Ribosomal, 16S - genetics
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Summary:The polymerase chain reaction (PCR) was used selectively to amplify specific rDNA sequences of Carnobacterium divergens, C. mobile, C. piscicola and C. gallinarum in purified DNA extracts, crude cell lysates and food samples. The PCR products were visualized by agarose gel electrophoresis and identified, at species level, by hybridization reactions with three specific oligonucleotide probes for C. divergens, C. mobile and C. piscicola/C. gallinarum designed from 16S rRNA sequence data. The PCR was sufficiently sensitive to amplify DNA from a single bacterium to detectable levels after 30 cycles of amplification. Both radioactive (32P) and non-radioactive alkaline phosphatase labelled probes was able to detect the PCR products. Detection was highly specific and the probes did not hybridize with DNA samples from any other of the bacterial species tested. These methods enabled the rapid and specific detection and identification of carnobacteria from pure cultures and samples of meat.
ISSN:0021-8847
2056-5232
DOI:10.1111/j.1365-2672.1992.tb01838.x