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Objective To develop a rapid method for detecting Salmonella, Escherichia coli O157, Listeria monocytogenes, Staphylococcus aureus, and Vibrio parahaemolyticus using multiplex polymerase chain reaction (PCR) and capillary electrophoresis technology. Methods Specific multiplex PCR primers were designed using conserved pathogenic genes of 5 kinds of foodborne pathogens, and the products were analyzed using capillary electrophoresis imaging. By optimizing the PCR reaction and electrophoresis conditions, a multiplex PCR capillary electrophoresis detection method was established. The specificity and sensitivity were studied as well. Results The specificity of the 5 pairs of primers was acceptable and no non-specific amplification occurred. The optimal primer concentration for multiplex PCR reaction was 0.2 μmol/L, and the optimal annealing temperature was 56.0 ℃. The optimal separation mode for capillary electrophoresis was AM900. The method had a high sensitivity, and the detection sensitivity could reach 5×10-3 ng/μL. Conclusion The multiplex PCR capillary electrophoresis method for detecting 5 kinds of foodborne pathogens established in this study is simple for operation with high specificity and sensitivity, and can be very practical in detection of foodborne pathogens.
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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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