Servizio Comunitario di Informazione in materia di Ricerca e Sviluppo - CORDIS

Pcr method for detection of histamine producer lactic acid bacteria in wine

Histamine is the most frequently biogenic amine (BA) involved in food intoxications. It is usually present at a low concentration in wine, but its toxic effect is strengthened by the presence of alcohol and other BAs of wine. Although there is no rule, some countries recommend upper limits of histamine ranging from 2 to 10mg per litre.

Histamine is produced by lactic acid bacteria (LAB) containing a specific enzyme, a histidine decarboxylase. Genes encoding the histidine decarboxylase of several LAB were described recently. Based on these gene sequences we have designed PCR primers which are useful for detecting histamine-producing LAB by quantitative PCR. In addition, we have optimised an existing procedure for the extraction of microbial DNA from wine samples.

The combination of both, the DNA extraction procedure and the quantitative PCR, allows for the detection and quantification of histamine-producing LAB in wine. Results obtained by this method were confirmed by other methods: plate counting, Southern blotting, quantitative PCR specific for another LAB gene, and HPLC determination of produced histamine.

This method appeared to be very accurate and sensitive, since it can detected as few as 2 histamine-producing cells per millilitre of wine and the quantification is precise up to 2x10e7 histamine-producing cells per millilitre. This method requires less than 8 hours to be performed and it can be carried out for many samples simultaneously. It is useful to detect histamine-producing LAB in must and wine at all stages of the winemaking in order to avoid histamine spoilage of wine.

Reported by

Faculty of Enology of Bordeaux 2 University
351, Cours de la Libération
33405 Talence
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