Coton Emmanuel, Coton Monika
ADRIA Normandie, Boulevard du 13 juin 1944, 14310 Villers-Bocage, France.
J Microbiol Methods. 2005 Dec;63(3):296-304. doi: 10.1016/j.mimet.2005.04.001. Epub 2005 Jun 1.
Formation of biogenic amines (BA) may occur in fermented foods and beverages due to the amino acid decarboxylase activities of Gram-positive bacteria. These compounds may cause food poisoning and therefore could imply food exportation problems. A set of consensual primers based on histidine decarboxylase gene (hdc) sequences of different bacteria was designed for the detection of histamine-producing Gram-positive bacteria. A multiplex PCR based on these hdc primers and recently designed primers targeting the tyrosine decarboxylase (tyrdc) gene was created. A third set of primers targeting the 16S rRNA gene of eubacteria was also used as an internal control. This multiplex PCR was performed on extracted DNA as well as directly on cell colonies. The results obtained show that this new molecular tool allowed for the detection of Gram-positive histamine- and/or tyramine-producing bacteria. The use of this molecular tool for early and rapid detection of Gram-positive BA-producing bacteria is of interest in evaluating the potential of cultured indigenous strains to produce biogenic amines in a fermented food product as well as to validate the innocuity of potential starter strains in the food industry.
由于革兰氏阳性菌的氨基酸脱羧酶活性，发酵食品和饮料中可能会形成生物胺（BA）。这些化合物可能会导致食物中毒，因此可能意味着食品出口问题。基于不同细菌的组氨酸脱羧酶基因（hdc）序列设计了一组共识引物，用于检测产生组胺的革兰氏阳性菌。基于这些hdc引物和最近设计的靶向酪氨酸脱羧酶（tyrdc）基因的引物创建了多重PCR。还使用了一组靶向真细菌16S rRNA基因的引物作为内部对照。这种多重PCR既在提取的DNA上进行，也直接在细胞菌落上进行。获得的结果表明，这种新的分子工具能够检测出产生组胺和/或酪胺的革兰氏阳性菌。使用这种分子工具对产生生物胺的革兰氏阳性菌进行早期快速检测，对于评估培养的本土菌株在发酵食品中产生生物胺的潜力以及验证食品工业中潜在发酵剂菌株的无害性具有重要意义。
