Detection of species-specific gene thermonuclease (nuc) in Staphylococcus aureus from chickens by real- time PCR assay.
BVMJ • 2014
Publication Information
Authors
Ashraf, A. AbdEl-Tawwab 1; Hanan, A. Fahmy 2; Fatma, I. El Hofy 1 and Abdelhalim, A.
Elshawadfy3
Keywords
Staphylococcus aureus, nuc gene, Real time PCR, SYBR Green
Journal
BVMJ
Publisher
Faculty of Vet.Med. Benha Univ.
Volume
27
Issue
2
Pages
415-427
publication.type
International
Paper Link
Not Available
Supplementary Materials
Not Available
Abstract
Accurate and rapid detection of S. aureus is crucial to control its infections and minimize its leakage to
the food chain. Therefore, the primary purpose of this research is to isolate S. aureus and characterize
the isolates by coagulase production from cloacal, tracheal and nasal swabs of apparently healthy broiler
& layer chickens and from different organs (joints, liver, lungs, ovaries, foot abscesses and heart blood)
of diseased broiler & layer chickens. A total of 318 samples were collected as follows: 108 sample from
different farms of apparently healthy broiler & layer and 210 samples from diseased layer & broiler
chickens. The samples were examined bacteriologically, 164 (51.6%) Staphylococcal isolates were
recovered from the 318 sample. 37/164 (22.6%) coagulase positive S. aureus were isolated. Upon
confirmation by Real Time PCR for thermonuclease (nuc) gene using SYBR Green, isolates showed
amplification at CT value 19, 20 and 21 with Melting temperature (Tm) values in the range of 76–77 oC.
Dissociation curve between 75-80oC which confirmed as S. aureus. The isolated S. aureus was highly
sensitive to vancomycin, amoxicillin + clavulinic acid and cephalothin by 84.5%, 83.8% and 78.4
respectively and highly resistant to ampicillin, oxacillin and penicillin by 75.7%, 73% and 70.2%
respectively
the food chain. Therefore, the primary purpose of this research is to isolate S. aureus and characterize
the isolates by coagulase production from cloacal, tracheal and nasal swabs of apparently healthy broiler
& layer chickens and from different organs (joints, liver, lungs, ovaries, foot abscesses and heart blood)
of diseased broiler & layer chickens. A total of 318 samples were collected as follows: 108 sample from
different farms of apparently healthy broiler & layer and 210 samples from diseased layer & broiler
chickens. The samples were examined bacteriologically, 164 (51.6%) Staphylococcal isolates were
recovered from the 318 sample. 37/164 (22.6%) coagulase positive S. aureus were isolated. Upon
confirmation by Real Time PCR for thermonuclease (nuc) gene using SYBR Green, isolates showed
amplification at CT value 19, 20 and 21 with Melting temperature (Tm) values in the range of 76–77 oC.
Dissociation curve between 75-80oC which confirmed as S. aureus. The isolated S. aureus was highly
sensitive to vancomycin, amoxicillin + clavulinic acid and cephalothin by 84.5%, 83.8% and 78.4
respectively and highly resistant to ampicillin, oxacillin and penicillin by 75.7%, 73% and 70.2%
respectively
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