Full
Outline
To establish a duplex real-time fluorescent polymerase chain reaction (RT-PCR) method for detecting Streptococcus salivarius subspecies and Lactobacillus delbrueckii subspecies bulgaricus in non-viable fermented milk.
The optimal reaction system of duplex RT-PCR was optimized by adjusting the ratio of 2 target primers/probes, primer/probe concentrations, Mg2+ concentration and annealing temperature. The specificity and reproducibility of the method were validated. Finally, the target bacteria were detected in the actual samples purchased from the market.
Optimal amplification was achieved in a 20 µL reaction system containing primer-probes for both targets at a 1:1 ratio, 0.3 µL each of forward and reverse primers (10 µmol/L), 0.15 µL each of probes (10 µmol/L), 3 mmol/L Mg2+ and an annealing temperature of 60 ℃. Both Streptococcus salivarius subspecies and Lactobacillus delbrueckii subspecies bulgaricus were detected in all 24 batches of commercial samples.
The established method enables rapid and accurate detection of Streptococcus salivarius subspecies and Lactobacillus delbrueckii subspecies bulgaricus in non-viable fermented milk, providing effective technical support for food authenticity supervision.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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