Identification of viable Lactobacillus cells in fermented dairy products

journal article - other

English

The combination PCR and ethidium monoazide (EMA-PCR) is a new metod for selective distinction between viable and dead bacterial cells. The general application of EMA is based on penetration into 'dead' cells with compromised membráně (cell wall systems) integrity. DNA bound to EMA cannot be PCR amplified. Live cells remain intact and only DNA from viable cells can be amplified. Experiments were performed using probiotic bacterial strain Lactobacillus paracasei subsp. paracasei CCDM 211/06 of human origin. The experimental conditions (time of cell incubation with EMA, and cell irradiation) were optimized. In the next step instead for live-dead cell distinction propidium monoazide was used. The Lactobacillus species specific primers were used for DNA amplification. The PCR products (250 bp) were separated using electrophoresis in 1.8 % agarose gel. The method was applied for live and dead cells identification in milk product (yoghurt).

ethidium monoazide, viable and dead bacterial cells

TRACHTOVÁ, Š.; ŠPANOVÁ, A.; RITTICH, B.

2008

9. 9. 2008

Chemické listy

Brno

1213-7103

Chemické listy

102

15

Czech Republic

805

806

2