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Development of specific and rapid detection of bacterial pathogens in dairy products by PCR

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Abstract

A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS),Staphylococcus aureus andEscherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on polymerase chain reaction (PCR) using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates, described previously only in human GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all knownsip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed withS. aureus andE. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/µL (i.e. 6000 CFU/mL) for GBS andE. coli, and 16 CFU/µL forS. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.

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Authors and Affiliations

  1. Institute of Molecular Biology, Slovak Academy of Sciences, 845 51, Bratislava, Slovakia

    M. Chotár & A. Godány

  2. Department of Nutrition and Food Assessment. Faculty of Chemical and Food Technology, Slovak Technical University, 812 37, Brotislava, Slovakia

    B. Vidová

Authors
  1. M. Chotár
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  2. B. Vidová
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  3. A. Godány
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Correspondence to A. Godány.

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This research was supported by projects of theMinistry of Agriculture of Slovak Republic (2003 SP 27/028, OE 02/028, OE 02-01-03).

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Chotár, M., Vidová, B. & Godány, A. Development of specific and rapid detection of bacterial pathogens in dairy products by PCR. Folia Microbiol 51, 639–646 (2006). https://doi.org/10.1007/BF02931632

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  • DOI: https://doi.org/10.1007/BF02931632

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