Propidium monoazide (PMA) was combined with real-time fluorescent quantitative PCR to detect DNA of living bacteria in milk products. A new, fast, and accurate method to detect Lactobacillus plantarum P-8 in fermented dairy products was developed. Influencing factors, such as concentrations, dark incubation, duration of illumination were determined to establish the optimal PMA treatment scheme. The results showed that after treating L. plantarum P-8 at 80℃ for 60 s, they became membrane-damaging. When 40 ug/mL PMA was used for 10 min dark incubation, followed by espousing for 20 min, PMA did not influence the living bacteria's DNA amplification. In addition, it inhibited DNA amplification of dead cells. A standard curve with a good linear relationship was set up based on L. plantarum P-8 plasmid standards. The relative coefficient of the curve was 0.992 9, and the lowest detection limit was 103 CFU/mL with good specificity. This method laid a foundation for improving the detection of viable probiotics in fermented milk products.
CRHAN Zhihao
,
GUO Shuai
,
HUANG Tian
,
ZHENG Yan
,
WANG Yuejiao
,
BAI Mei
,
WANG Jicheng ZHANG Heping
. Detection of viable Lactobacillus plantarum P-8 in fermented milk using propidium monoazide combined with real-time fluorescence quantitative P[J]. Food and Fermentation Industries, 2019
, 45(1)
: 183
-189
.
DOI: 10.13995/j.cnki.11-1802/ts.018232
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