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Simultaneous detection of Listeria monocytogenes and Salmonella spp. in dairy products using real time PCR-melt curve analysis

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Abstract

The present investigation reports development of post real time PCR (RTi-PCR) - melt curve analysis for simultaneous detection of Listeria monocytogenes and Salmonella spp. The optimal Sybr Green I (SG-I) concentration of 1.6 μM resulted in two specific peaks with melting temperature (Tm) of 79.90 ± 0.39 °C and 86.29 ± 0.13 °C for L. monocytogenes and Salmonella spp respectively. The detection sensitivity of the assay in reconstituted non-fat dried milk (NFDM; 11%) spiked with the target pathogens at different levels was 3 log cfu per ml of each pathogen. However, the sensitivity was improved up to 1 log cfu per ml by including pre-enrichment step of 6 h. On application of assay on 60 market samples, one sample each of raw milk and ice cream was detected positive for L. monocytogenes and Salmonella spp. Assay was quite specific as no cross reactivity with non target cultures could be observed. The developed assay can find valuable application in monitoring dairy products for the presence of L. monocytogenes and Salmonella spp. to ensure their microbiological quality and safety.

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Acknowledgement

The authors gratefully acknowledge the financial assistance provided by the Ministry of Food Processing Industry, Govt. of India, for the procurement of Real Time PCR instrument. Jitender Singh duly acknowledges the financial assistance from Indian Council of Agricultural Research and facilities provided by Director, NDRI, Karnal.

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  1. Jitender Singh

    Present address: Product and Process Development Group, CALF Building, National Dairy Development Board, Anand, 388001, Gujarat, India

Authors and Affiliations

  1. Molecular Biology Unit, Dairy Microbiology Division, National Dairy Research Institute, Karnal, 132001, Haryana, India

    Jitender Singh, Virender K. Batish & Sunita Grover

Authors
  1. Jitender Singh
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  2. Virender K. Batish
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  3. Sunita Grover
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Correspondence to Sunita Grover.

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Singh, J., Batish, V.K. & Grover, S. Simultaneous detection of Listeria monocytogenes and Salmonella spp. in dairy products using real time PCR-melt curve analysis. J Food Sci Technol 49, 234–239 (2012). https://doi.org/10.1007/s13197-011-0278-3

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  • DOI: https://doi.org/10.1007/s13197-011-0278-3

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