Rapid detection of Salmonella and Listeria monocytogenes in milk by immunomagnetic separation and polymerase chain reaction

A rapid detection method combining immunomagnetic separation (IMS), PCR and slot blot detection was developed for the detection of Salmonella and Listeria monocytogenes in milk. Bacteria were first isolated and concentrated from phosphate-buffered saline (PBS) or milk by IMS. After extraction from diluted bacteria culture with the extraction buffer, bacterial DNA was subjected to PCR. Slot blot assay was optimized and used to measure PCR products. The lowest level of detection by this method was 40 cfu/ml in PBS or milk for both pathogens. The whole detection procedure could be completed within 7 h. Moreover, this detection method is simple and easy to handle for a large number of samples. Using multiplex PCR (amplification of two different bacterial DNA in the same PCR tube) and slot blot, simultaneous detection of both bacteria was also assessed. The detection sensitivities of 103 cfu/ml for both bacteria were the same as when PCR and slot blot were used for each bacterium separately. The combination of IMS, PCR and slot blot seems to give a highly sensitive and time-efficient procedure, which could be used for routine detection of Salmonella and L. monocytogenes in milk