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Development and evaluation of two novel oligonucleotide probes based on 16S rRNA sequence for the identification of Salmonella in foods
Author(s) -
Lin C.K.,
Tsen H.Y.
Publication year - 1995
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1995.tb03093.x
Subject(s) - citrobacter , salmonella , 16s ribosomal rna , microbiology and biotechnology , biology , enterobacteriaceae , serratia marcescens , genbank , serratia , oligomer restriction , klebsiella pneumoniae , proteus vulgaris , escherichia coli , bacteria , oligonucleotide , dna , gene , genetics , pseudomonas
DNA sequence in the V3 to V6 region of the 16S rRNA gene of Salmonella enteritidis was determined. By comparison of this sequence with those of Escherichia coli and Proteus vulgaris obtained from GenBank/EMBL database, three oligonucleotides termed as 16S I, 16S II and 16S III were synthesized. Hybridization of these oligonucleotides with 325 Salmonella isolates and some non‐ Salmonella isolates including the Salmonella closely related species of the family of Enterobacteriaceae showed that 16S II could not be used as a Salmonella specific‐probe. 16S I and 16S III hybridized with all the Salmonella isolates tested, the former also hybridizing with Citrobacter spp. and the latter hybridizing with Klebsiella pneumoniae as well as Serratia marcescens. Since enrichment of the target cells in food samples was usually required prior to the DNA hybridization assay, the interference from those non‐ Salmonella isolates could be prevented by enrichment by culturing in lactose‐combined tetrathionate (CTET) broth followed by Gram‐negative (GN) broth at 37°C and/or 43°C. Such a culture step could inhibit the growth of Klebsiella spp., Ser. marcescens and/or Citrobacter spp. and allowed the specific detection of Salmonella .