Comparison of conventional and molecular methods for the routine confirmation of Listeria monocytogenes in milk products produced domestically in Croatia

Abstract

Conventional methods for the identification of Listeria in foodstuffs are generally cumbersome and time consuming. The use of primary enrichment in half strength Fraser broth and the use of PALCAM agar were assessed in comparison with API Listeria and polymerase chain reaction (PCR) for their ability to accurately detect and confirm the presence of List. monocytogenes in milk products. The aim of our work was to detect List. monocytogenes in domestic unpasteurised milk, fresh cheese and cream of raw milk taken from four different district of Zagreb-Croatia using conventional (microbiological and biochemical - API test) and PCR methods. Of the 180 milk products samples tested, 27.6% were presumptively positive for Listeria on PALCAM agar. Only 21.3% of samples were confirmed to be positive for Listeria by API Listeria test, and 17.3% were confirmed to be positive for List. monocytogenes by PCR amplification of the hly gene (64 bp). PCR was able to eliminate the false positive and detect all List. monocytogenes in the milk products, unlike the conventional methods used in the industry. These results indicate a low presence of this pathogen in this area (Zagreb) of Croatia. PCR proves to be a sensitive and rapid technique to be included in the procedure of detection of List. monocytogenes in food products and this method is considerably faster than current standard methods.