Species-specific real-time PCR assay for enumeration of Anoxybacillus flavithermus and Geobacillus stearothermophilus spores in dairy products
| Autor: | Yoshiaki Ohkubo, Hidemasa Motoshima, Naoya Katano, Kenji Uchida |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
food.ingredient
biology Chemistry Thermophile fungi 0402 animal and dairy science food and beverages 04 agricultural and veterinary sciences Proteinase K biology.organism_classification 040401 food science 040201 dairy & animal science Applied Microbiology and Biotechnology Spore chemistry.chemical_compound 0404 agricultural biotechnology food Skimmed milk biology.protein Enumeration SYBR Green I Geobacillus stearothermophilus Food science Bacteria Food Science |
| Zdroj: | International Dairy Journal. 93:15-21 |
| ISSN: | 0958-6946 |
| DOI: | 10.1016/j.idairyj.2019.01.015 |
| Popis: | The thermophilic spore-forming bacteria, Anoxybacillus flavithermus and Geobacillus stearothermophilus, grow readily, especially on milk-powder processing lines, and are important for processing-plant hygiene management. We developed a real-time PCR assay using SYBR Green I to monitor the spores of these species in dairy products. We designed new primer pairs specific for each species from the nucleotide sequences of the stage 0 sporulation gene A (spo0A). Ethidium monoazide treatment enabled us to quantify only spores. In dairy products with high protein content, treatment with proteinase K allowed precise quantification of spores. This assay was superior for counting spores, having high linearity (r2 = 0.99) and a wide quantification range (101 to 106 spores mL−1). This new method can be applied to quantification of spores in samples of milk, skimmed milk, and cream. |
| Databáze: | OpenAIRE |
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