The design and optimisation of the Multiplex PCR for detection of <i>Mycoplasma agalactiae</i>, <i>Yersinia enterocolitica</i> and <i>Aeromonas hydrophila</i> in raw milk
MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL, cilt.56, sa.7, ss.377-380, 2001 (SCI-Expanded)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 56 Sayı: 7
- Basım Tarihi: 2001
- Dergi Adı: MILCHWISSENSCHAFT-MILK SCIENCE INTERNATIONAL
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED)
- Sayfa Sayıları: ss.377-380
- Hacettepe Üniversitesi Adresli: Evet
Özet
Single-locus (sl) and multiplex (m) polymerase chain reaction (PCR) procedures were developed for the detection of M. agalactiae, Y. enterocolitica and A. hydrophila in raw milk samples. Three oligonucleotide primers for each pathogen were used in PCR, to detect a 375 base-pair (bp) fragment of M. agalactiae chromosomal DNA, the yst gene (145 bp) of Y. enterocolitica and aer gene (209 bp) of A. hydrophila. In optimisation studies, the sensitivity limits of sIPCR were found 10.0, 5.0 and 1.0 pg for M. agalactiae, Y. enterocolitica and A. hydrophila, respectively. 2.0 mM MgCl2 concentration was found as superior for mPCR. When studied with naturally contaminated raw milk samples, detection rates obtained by PCR were 1.8%, 53% and 23% for M. agalactiae, Y. enterocolitica and A. hydrophila, respectively. Results indicate the ability to detect these th ree pathogens effectively on the basis of direct PCR analysis of raw milk.