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

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...

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Bibliographic Details
Published in:Journal of dairy science 2002-07, Vol.85 (7), p.1717-1723
Main Authors: MEIRI-BENDEK, I, LIPKIN, E, FRIEDMANN, A, LEITNER, G, SARAN, A, FRIEDMAN, S, KASHI, Y
Format: Article
Language:English
Subjects:
Animal bacterial diseases
Animal productions
Animals
Bacterial diseases
Base Sequence
Biological and medical sciences
Cattle
DNA Primers
DNA, Bacterial - analysis
DNA, Mitochondrial - analysis
Female
Fundamental and applied biological sciences. Psychology
Infectious diseases
Mastitis, Bovine - diagnosis
Mastitis, Bovine - microbiology
Medical sciences
Milk - microbiology
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - veterinary
RNA, Ribosomal, 16S - genetics
Sensitivity and Specificity
Streptococcal Infections - diagnosis
Streptococcal Infections - veterinary
Streptococcus agalactiae - genetics
Streptococcus agalactiae - isolation & purification
Terrestrial animal productions
Time Factors
Vertebrates
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Summary: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.
ISSN:0022-0302
1525-3198
DOI:10.3168/jds.S0022-0302(02)74245-8