Development of a quantitative molecular method for enumeration of Salmonella enterica

Authors

  • Marija TRKOV Univ. of Ljubljana, Biotechnical Fac., Zootechnical Dept., Groblje 3, SI-1230 Domžale, Slovenia
  • Gorazd AVGUŠTIN Univ. of Ljubljana, Biotechnical Fac., Zootechnical Dept., Groblje 3, SI-1230 Domžale, Slovenia

DOI:

https://doi.org/10.14720/aas.2002.80.2.15576

Keywords:

microbiology, bacteria, Salmonella enterica, molecular genetics, methods, c-PCR, capillary electrophoresis

Abstract

A competitive polymerase chain reaction (c-PCR) was developed for rapid enumeration of Salmonella enterica cells in broth culture. The primers MINF and MINR-Hex ensured the methods specificity. An internal DNA control was prepared by enzymatic restriction and subsequent ligation of the flanking regions of the amplified 16S rRNA gene of S. Agona (U92197) strain. The prepared internal control had the same primer specific sequences as the target DNA but was 27 bp shorter. The c-PCR products were quantified by capillary electrophoresis and the results were obtained by regression analysis. Using the described system and 5 × 10–5 times diluted internal control, a quantification of S. enterica cells in broth culture in the range between 1.2 × 104 and 6 × 105 per reaction mixture was made possible.   The preliminary test showed that the results of the c-PCR quantification of the salmonella cells in broth culture are 2.9 times higher in comparison with the results of the indirect counting method. The authors are still convinced, however, that the method offers a rapid, specific and sufficiently accurate counting of S. enterica cells in natural microbiological samples.

Published

15. 12. 2002

Issue

Section

Original Scientific Article

How to Cite

TRKOV, M., & AVGUŠTIN, G. (2002). Development of a quantitative molecular method for enumeration of Salmonella enterica. Acta Agriculturae Slovenica, 80(2), 147–155. https://doi.org/10.14720/aas.2002.80.2.15576

Similar Articles

1-10 of 378

You may also start an advanced similarity search for this article.

Most read articles by the same author(s)

1 2 > >>