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  • Küçük Resim
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    Development of pmp Gene-Specific PCR Assay with A Host Specific Internal Control for Chlamydophila felis
    (Chulalongkorn Univ, 2014) Cantekin, Zafer; Solmaz, Hasan; Altug, Nuri; Ozmen, Gamze Ozge
    Chlamydophila felis is the primary bacterial agent of conjunctivitis and upper respiratory disease in cats. Transmission of the disease requires close contact between cats. Polymerase chain reaction is a useful tool for detection of this organism. The aim of this study was to develop a polymerase chain reaction assay with an internal amplification control for the detection of C. felis. Primer pairs were designed specifically for polymorphic membrane protein gene of C. felis and cytochrome b gene of cat, and their specificity and sensitivity were examined. Primers specific for both genes were then multiplexed. In the simplex polymerase chain reaction analyses with 10-fold dilutions, C. felis DNA was detected with designed primers for polymorphic membrane protein genes up to 1.6 pg/mu l and cat DNA was demonstrated in all samples in the polymerase chain reaction. Moreover, in the multiplex polymerase chain reaction, C. felis DNA and cat DNA were detected together. These designed primers specific for C. felis might have potential for research on infections and shedding of this organism in cats as the internal control host specific primers might have potential for using internal control for detection of different microorganisms in cats.
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    Öğe
    Development of pmp gene-specific PCR assay with a host specific internal control for Chlamydophila felis
    (Chulalongkorn University Printing House, 2014) Cantekin, Zafer; Solmaz, Hasan; Altug, Nuri; Ozmen, Gamze Ozge
    Chlamydophila felis is the primary bacterial agent of conjunctivitis and upper respiratory disease in cats. Transmission of the disease requires close contact between cats. Polymerase chain reaction is a useful tool for detection of this organism. The aim of this study was to develop a polymerase chain reaction assay with an internal amplification control for the detection of C. felis. Primer pairs were designed specifically for polymorphic membrane protein gene of C. felis and cytochrome b gene of cat, and their specificity and sensitivity were examined. Primers specific for both genes were then multiplexed. In the simplex polymerase chain reaction analyses with 10-fold dilutions, C. felis DNA was detected with designed primers for polymorphic membrane protein genes up to 1.6 pg/?l and cat DNA was demonstrated in all samples in the polymerase chain reaction. Moreover, in the multiplex polymerase chain reaction, C. felis DNA and cat DNA were detected together. These designed primers specific for C. felis might have potential for research on infections and shedding of this organism in cats as the internal control host specific primers might have potential for using internal control for detection of different microorganisms in cats.
  • Küçük Resim
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    PCR ASSAY WITH HOST SPECIFIC INTERNAL CONTROL FOR STAPHYLOCOCCUS AUREUS FROM BOVINE MILK SAMPLES
    (Univ Sv Kiril & Metodij Skopje, Fak Veterinarna Medicina, 2015) Cantekin, Zafer; Ergun, Yasar; Solmaz, Hasan; Ozmen, Gamze Ozge; Demir, Melek; Saidi, Radhwane
    Staphylococcus aureus is considered as one of the most important and common pathogens of bovine mastitis. Polymerase Chain Reaction is frequently proposed in the diagnosis of S. aureus directly from milk samples instead of classical culture. However, false-negative results may occur in the polymerase chain reaction analysis performed directly from clinical material. For the purpose of disclosing the false negative results, the use of internal amplification controls can be beneficial. Therefore, in this study a new polymerase chain reaction technique with host specific internal amplification control was developed by optimizing S. aureus-specific primers in combination with bovine specific primers. The effectiveness of the developed technique in this study was attempted in milk samples from bovine subclinical mastitis. This technique has the potential to detect S. aureus from bovine milk samples or dairy products.
  • Küçük Resim
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    Presence of Antibiotic Resistance Genes in Staphylococci Isolated From Bovine Subclinical Mastitis
    (Sivar-Soc Italiana Veterinari Animali Reddito, 2024) Cantekin, Zafer; Ozmen, Gamze Ozge; Demir, Melek; Er, Zeynep Yilmaz; Gurturk, Kemal; Solmaz, Hasan; Ekin, Ismail Hakki
    The prevalence of antibiotic resistance increases rapidly worldwide, and the primary culprit is represented by their widespread use. Subclinical mastitis is the leading cause of most antibiotic treatment, representing also one of the significant problems of bovine herd management. One of the main causes of subclinical mastitis is Staphylococcus aureus. Therefore, the determination of antibiotic resistance against Staphylococcus aureus is an essential step in the treatment of subclinical mastitis. The aim of this study was to identify antibiotic resistance genes in staphylococci obtained from cases of bovine subclinical mastitis in three provinces and the relationship between antibiotic resistance and ease of antibiotic availability (Burdur, Hatay and Van) in Turkey. In total, 283 isolates (Burdur, n = 36; Hatay, n = 47; Van, n = 200 isolates) were studied. The isolates were first identified as Staphylococcus aureus and/or non-aureus staphylococci (NAS) by conventional phenotypic methods, and the species was then confirmed by a multiplex polymerase chain reaction (PCR). A simplex PCR assay was performed to detect antibiotic resistance genes (mecA, mecC, aacA-aphD, ermA, ermB, ermC, tetK, tetM and blaZ). Among the isolates from all three provinces, the blaZ gene was the most prevalent antibiotic resistance gene, present in 43 out of 156 (28%) NAS isolates, 27 out of 127 (21%) S. aureus isolates and 25% of all the isolates. In contrast, tetM was the most prevalent gene in the Hatay isolates, detected in 64% of all isolates. The mecA-gene was present in 10% of the NAS, and in 3% of the S. aureus isolates. The mecC and ermA genes were not detected in any of the isolates. This shows that antimicrobial resistance, as determined by PCR, is common in Staphylococcus isolates from mastitis in Turkey, and warrants systematic treatment protocols as well as the implementation of preventative strategies to reduce antimicrobial usage.