Determinasi Genetik Staphylococcus aureus Sapi Perah di Baturraden dan Pengembangan Deteksi Stafilokokal Mastitis Langsung dari Susu Segar dengan Polymerase Chain Reaction (PCR)
| Main Authors: | , FATKHANUDDIN AZIZ, , Prof. Dr. drh. Siti Isrina Oktavia Salasia, |
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| Format: | Thesis NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2013
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/119498/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=59501 |
Daftar Isi:
- Staphylococcus aureus (S. aureus) is one of bacteria that cause subclinical mastitis with significant economic losses in dairy farm. Staphylococcus aureus could also cause bacterial foodborne disease. Identification of the pathogenic bacteria in raw milk can be used for a definitive diagnosis of food poisoning source, it can be used as an important information for preventive and control the source of food poisoning. This research aimed to determine the genetic profiles of S. aureus and to develop the detection of S. aureus directly from raw milk by PCR to get a rapid and accurate method for screening the quality of milk. In this study used 50 samples of raw milk collected from dairy cows in Baturraden. Staphylococcus aureus were identified conventionally based on bacterial culture assays, mannitol salt agar (MSA), koagulase, catalase, VP tests and Gram staining. Staphylococcus aureus were further identified by PCR to determine the 23S rRNA gene and the virulence determinants encoded by coa, nuc, clfA, fnbA, fnbB, cap5, spa IgG and spa X- region genes. The clonal relationship among isolates was carried out based on AFLP analysis through calculation unweight the pair group method with arithmatic average (UPGMA) and compared with S. aureus isolated from other regions. Development of detection of S. aureus directly from raw milk was done by extraction of DNA from raw milk samples, and then analyzed by PCR using primer species specific for 23S rRNA gene detection. The specificity and sensitivity of the detection of S. aureus directly from the raw milk compared with the results of detection of S. aureus through bacterial cultrures conventionally and molecularly. The research resulted 17 isolates positive for S. aureus, could be determined the virulence determinant genes encoded clfA, coa, spa IgG and spa X- region (100%), cap5 gene (88%) and fnbB gene (65%). AFLP analysis showed that all isolates of S. aureus originated from Baturraden could be grouped in 1 cluster with similaritas coefficients of 80%. It was of interest that S. aureus strain BR49 2 and BR49 3 were detected in a close relationship (similarity coefficients of 100%), had also with the size of the coa gene (600bp), spa X-region gene (100bp) and both of strains did not have of cap5 gene. Staphylococcus aureus could be directly identified from raw milk with DNA extraction methods and by PCR analysis of 23S rRNA gene. The results showed that there were no difference between the direct method for detection of S. aureus from raw milk with the detection from bacterial cultures.