Multiplex PCR method for the simultaneous detection of histamine-, tyramine-, and putrescine-producing lactic acid bacteria in foods.
In a screening of primers, we have selected three pairs of primers for a multiplex PCR assay for the simultaneous detection of lactic acid bacteria (LAB) strains, which potentially produce histamine, tyramine, and putrescine on fermented foods. These primers were based on sequences from histidine, tyrosine, and ornithine decarboxylases from LAB. Under the optimized conditions, the assay yielded a 367-bp DNA fragment from histidine decarboxylases, a 924-bp fragment from tyrosine decarboxylases, and a 1,446-bp fragment from ornithine decarboxylases. When the DNAs of several target organisms were included in the same reaction, two or three corresponding amplicons of different sizes were observed. This assay was useful for the detection of amine-producing bacteria in control collection strains and in a LAB collection. No amplification was observed with DNA from nonproducing LAB strains. This article is the first describing a multiplex PCR approach for the simultaneous detection of potentially amine-producing LAB in foods. It can be easily incorporated into the routine screening for the accurate selection of starter LAB and in food control laboratories.
Marcobal
Angela
A
Departamento Microbiología, Instituto de Fermentaciones Industriales, Consejo Superior de Investigaciones Científicas, 28006 Madrid, Spain.
de las Rivas
Blanca
B
Moreno-Arribas
M Victoria
MV
Muñoz
Rosario
R
D000595:Amino Acid Sequence; D015169:Colony Count, Microbial; D053938:DNA Fragmentation; D004269:DNA, Bacterial / Q000032:analysis*; D005516:Food Microbiology; D006632:Histamine / Q000737:chemistry; D007778:Lactobacillus / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods*; D011700:Putrescine / Q000737:chemistry; D012680:Sensitivity and Specificity; D014439:Tyramine / Q000737:chemistry
D004269:DNA, Bacterial; D006632:Histamine; D011700:Putrescine; D014439:Tyramine