A PCR-based method for the detection of Streptococcus agalactiae in milk

I. Meiri-Bendek, E. Lipkin, A. Friedmann, G. Leitner, A. Saran, S. Friedman, Y. Kashi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

69 Scopus citations

Abstract

Bovine mastitis caused by Streptococcus agalactiae is mainly subclinical and therefore can be diagnosed only in the laboratory. We developed a polymerase chain reaction (PCR)-based method for specific and sensitive detection of S. agalactiae in raw milk. The specificity of the PCR reaction is based on unique S. agalactiae DNA sequences within the 16S subunit of the rRNA genes. Two pairs of sequences were used as positive controls; general streptococci primers, which anneal to conserved areas within the 16S rRNA subunit gene, and primers, which anneal to sequences within bovine mitochondrial DNA. The method of detection includes selective enrichment of S. agalactiae in the milk sample, followed by DNA extraction using a rapid and simple procedure developed for this purpose, and specific PCR reaction with appropriate controls. The method enables the detection of one bacterium in 1 ml of raw milk. The method developed can be easily incorporated as part of routine screening of bulk milk collection tanks for early detection of infected cows in a herd.

Original languageEnglish
Pages (from-to)1717-1723
Number of pages7
JournalJournal of Dairy Science
Volume85
Issue number7
DOIs
StatePublished - Jul 2002

Keywords

  • Bovine mastitis
  • Polymerase chain reaction diagnosis
  • Streptococcus agalactiae

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