PMID- 12694460
OWN - NLM
STAT- MEDLINE
DCOM- 20030729
LR  - 20190916
IS  - 1364-5072 (Print)
IS  - 1364-5072 (Linking)
VI  - 94
IP  - 5
DP  - 2003
TI  - Specific detection of Pseudomonas spp. in milk by fluorescence in situ 
      hybridization using ribosomal RNA directed probes.
PG  - 936-45
AB  - AIMS: Pseudomonas spp. are considered the most important milk spoilage organisms. 
      Here we describe development of a fluorescence in situ hybridization (FISH) probe 
      specific for detection and enumeration of Pseudomonas spp. in milk. METHODS AND 
      RESULTS: 16S rRNA sequences were analysed to develop specific oligonucleotide 
      probe for the genus Pseudomonas. Twenty different Pseudomonas spp. and 23 
      bacterial species from genera other than Pseudomonas (as negative controls) were 
      tested. All tested Pseudomonas spp. yielded a positive FISH reaction, whereas 
      negative controls showed no FISH reaction except for Burkholderia cepacia that 
      showed a relatively weak FISH reaction. The FISH assay specifically stains 
      Pseudomonas in milk when the milk contains a mixture of other bacterial species. 
      The FISH assay takes 2 h and compares favourably with current culturing methods, 
      which take a minimum of 48 h. Specificity of the probe was validated using 
      polymerase chain reaction to selectively amplifying the Pseudomonas rDNA gene and 
      sequencing the gene products. CONCLUSIONS: The method presented in this study 
      allows simultaneously detection, identification and enumeration of Pseudomonas 
      spp. in milk. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and accurate 
      enumeration of Pseudomonas facilitates the identification of specific 
      contamination sources in dairy plants, the accurate validation of pasteurization 
      treatments and the prediction of shelf life of processed milk.
FAU - Gunasekera, T S
AU  - Gunasekera TS
AD  - Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, 
      Australia. gunaseke@msu.edu
FAU - Dorsch, M R
AU  - Dorsch MR
FAU - Slade, M B
AU  - Slade MB
FAU - Veal, D A
AU  - Veal DA
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
RN  - 0 (DNA, Bacterial)
RN  - 0 (RNA Probes)
RN  - 0 (RNA, Bacterial)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Animals
MH  - Bacteriological Techniques/methods
MH  - DNA, Bacterial/genetics
MH  - Flow Cytometry/methods
MH  - *Food Microbiology
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Milk/*microbiology
MH  - Polymerase Chain Reaction/methods
MH  - Pseudomonas/genetics/*isolation & purification
MH  - RNA Probes
MH  - RNA, Bacterial/genetics
MH  - RNA, Ribosomal, 16S/genetics
MH  - Sensitivity and Specificity
EDAT- 2003/04/16 05:00
MHDA- 2003/07/30 05:00
CRDT- 2003/04/16 05:00
PHST- 2003/04/16 05:00 [pubmed]
PHST- 2003/07/30 05:00 [medline]
PHST- 2003/04/16 05:00 [entrez]
AID - 1930 [pii]
AID - 10.1046/j.1365-2672.2003.01930.x [doi]
PST - ppublish
SO  - J Appl Microbiol. 2003;94(5):936-45. doi: 10.1046/j.1365-2672.2003.01930.x.