Journal
FOOD CONTROL
Volume 57, Issue -, Pages 185-189Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.foodcont.2015.03.050
Keywords
Propidium monoazide; Taqman-based qPCR; Viable Salmonella; Raw shrimp
Categories
Funding
- National Natural Science Foundation of China [31271870]
- project of Science and Technology Commission of Shanghai Municipality [14DZ1205100, 14320502100, 12391901300]
- Key Project of Shanghai Agriculture Prosperity through Science and Technology [2014, 3-5]
- Cross-discipline project [B5201120040]
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Propidium monoazide (PMA) combined with molecular quantitative real-time PCR (qPCR) has been widely used for only detection of viable bacteria. However, recent studies indicated PMA did not fully inhibit the detection of dead Salmonella. In this study, we developed a more effective PMA Taqman-based qPCR than previous studies to quantify viable Salmonella spp. in raw shrimp. This method has high specificity by using 60 strains belonging to 23 species. The optimization of the PMA concentration showed that 100 mu M was considered optimal to effectively inhibit 10(6) CFU/mL dead cells, while only 10(3)-10(4) CFU/mL dead cells could be inhibited in previous reports. This assay could detect viable Salmonella spp. at as low as 36 CFU/mL in pure culture and 100 CFU/g in raw shrimp. By comparing with PMA-qPCR, qPCR and plate counting for quantifying Salmonella in samples, this PMA-qPCR was obviously superior to qPCR and had good agreement with plate counting. In conclusion, this effective method can be used as an available tool to quantify viable Salmonella spp. in raw shrimp. (C) 2015 Elsevier Ltd. All rights reserved.
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