Development of specific fluorescent oligonucleotide probes for in situ identification of wine lactic acid bacteria.
A rapid method for the identification of lactic acid bacteria (LAB) from wine has been developed. This method is based on fluorescence in situ hybridisation (FISH), using fluorescent oligonucleotide probes, homologous to 16S rDNA of those species of LAB commonly found in wines. The protocol for the specific detection of these bacteria was established through the hybridisation of 36 reference strains. The specificity of the probes was evaluated by using pure cultures. Probes were used to identify species in different wines, making it evident that direct identification and quantification from natural samples without culturing is also possible. The results show that FISH is a promising technique for the rapid identification of LAB, allowing positive identification in a few hours (4-16 h).
Blasco
Lucía
L
ENOLAB-Laboratori de Microbiologia Enològica, Departament de Microbiologia i Ecologia, Facultat de Ciències Biològiques, Universitat de València, Dr. Moliner 50, Burjassot, E-46100 Valencia, Spain.
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D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005285:Fermentation; D005456:Fluorescent Dyes / Q000302:isolation & purification; D017404:In Situ Hybridization, Fluorescence; D019344:Lactic Acid / Q000378:metabolism; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007934:Leuconostoc / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015336:Molecular Probe Techniques; D015345:Oligonucleotide Probes / Q000235:genetics / Q000302:isolation & purification; D010376:Pediococcus / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014920:Wine / Q000382:microbiology*
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D019344:Lactic Acid