TY - JOUR
T1 - A PCR-based method for the detection of Streptococcus agalactiae in milk
AU - Meiri-Bendek, I.
AU - Lipkin, E.
AU - Friedmann, A.
AU - Leitner, G.
AU - Saran, A.
AU - Friedman, S.
AU - Kashi, Y.
PY - 2002/7
Y1 - 2002/7
N2 - 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.
AB - 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.
KW - Bovine mastitis
KW - Polymerase chain reaction diagnosis
KW - Streptococcus agalactiae
UR - http://www.scopus.com/inward/record.url?scp=0036653678&partnerID=8YFLogxK
U2 - 10.3168/jds.S0022-0302(02)74245-8
DO - 10.3168/jds.S0022-0302(02)74245-8
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C2 - 12201522
AN - SCOPUS:0036653678
SN - 0022-0302
VL - 85
SP - 1717
EP - 1723
JO - Journal of Dairy Science
JF - Journal of Dairy Science
IS - 7
ER -