Yıl 2018, Cilt 21, Sayı 6, Sayfalar 957 - 965 2018-11-30

Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus
Staphylococcus aureus Klinik İzolatlarında Gentamisin, Penisilin, Metisilin, Vankomisin, Linezolid ve Tetrasiklin Direncinin Fenotipik ve Genotipik Olarak Araştırılması

Zerife ORHAN [1] , Arzu KAYIŞ [2] , Esra KAYA [3] , Murat ARAL [4]

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In this study, it was aimed to investigate the resistance rates of gentamicin, penicillin, methicillin, vancomycin, linezolid and tetracycline by phenotypic and genotypic methods in Staphylococcus aureus isolates and to determine plasmid content. Between the months of January and September in 2015, 100 clinical isolates of S. aureus were obtained from different samples such as wound, blood, urine. The automated bacteria identification and antibiotic susceptibility system (BD PhoenixTM, Sparks, MD, USA) was used to determine of antibiotic sensitivities. The resistance to methicillin was also investigated by Kirby-Bauer disc diffusion method using a 30 μg cefoxitin disc. The presence of aac(6’)/aph(2’’), blaZ, mecA, femA, vanA, vanB, cfr, tetK and tetM genes related to antibiotic resistance was investigated by PCR amplification in all isolates. Plasmid DNAs were isolated by using a Thermo Scientific GeneJET Plasmid Miniprep Kit. The cefoxitin resistance of S.aureus isolates, identified according to the results of disk diffusion and automated system, was calculated as 19%. Vancomycin and linezolid resistance were not observed in isolates while gentamicin 2%, penicillin 100%, methicillin 19%, tetracycline 18% resistance were identified using the automated system. According to the results of molecular analysis aac(6’)/aph(2’’), blaZ, mecA, femA, tetK and tetM genes frequencies were determined as 2%, 100%, 19%, 100%, 17% and 3% respectively, but vanA, vanB and  cfr genes were not amplified by PCR. In order to determine the relationship between antibiotic resistance and plasmid presence, plasmids were isolated from identified bacterial isolates. It is found that most of bacterial isolates (79%) contain different numbers plasmids. Rapid and reliable method for antibiotic susceptibility is important to determine the appropriate therapy decision. PCR can be used for confirmation of the results obtained by automated system or could be used as an alternative diagnostic method in the routine diagnosis for rapid, sensitive, and specific detection of MRSA associated antibiotic resistance genes.

Bu çalışmada, Staphylococcus aureus izolatlarında gentamisin, penisilin, metisilin, vankomisin, linezolid ve tetrasiklin direnç oranlarının araştırılması ve plazmid içeriğinin belirlenmesi amaçlanmıştır. 2015 yılı Ocak-Eylül ayları arasında yara, kan, idrar gibi farklı örneklerden 100 tane S. aureus izolatı elde edilmiştir. Antibiyotik duyarlılıklarını belirlemek için otomatik bakteri identifikasyon ve antibiyotik duyarlılık sistemi (BD PhoenixTM, Sparks, MD, ABD) kullanıldı. Ayrıca, metisilin direnci 30 μg sefoksitin disk kullanılarak Kirby-Bauer disk difüzyon yöntemi ile de araştırıldı. Antibiyotik direncine bağlı aac (6 ')/aph (2' '), blaZ, mecA, femA, vanA, vanB, cfr, tetK ve tetM genlerinin varlığı tüm izolatlarda PCR amplifikasyonu ile araştırıldı. Plazmid DNA'ları Thermo Scientific GeneJET Plazmid Miniprep Kiti kullanılarak izole edildi. Disk difüzyon ve otomatik sistem sonuçlarına göre belirlenen S.aureus izolatlarının sefoksitin direnci %19 olarak belirlendi. Vankomisin ve linezolid direnci izolatlarda görülmezken, otomotize sistem ile %2 gentamisin, %100 penisilin, %19 metisilin, %8 tetrasiklin direnci belirlendi. Moleküler analiz sonuçlarına göre aac (6 ')/aph (2' '), blaZ, mecA, femA, tetK ve tetM genleri sırasıyla % 2, % 100, % 19, % 100, % 17 ve %3 olarak belirlendi. Fakat vanA, vanB ve cfr genleri PCR ile amplifiye edilmedi. Antibiyotik direnci ve plazmid varlığı arasındaki ilişkiyi belirlemek için plazmidler, tanımlanmış bakteriyel izolatlardan izole edildi. Bakteriyel izolatların çoğunun (% 79) farklı sayılarda plazmid içerdiği bulundu.  

Antibiyotik duyarlılığı için hızlı ve güvenilir bir yöntem, uygun tedavi kararını belirlemek için önemlidir. Otomatik sistem ile elde edilen sonuçların doğrulanması için PCR kullanılabilir veya MRSA ile ilişkili antibiyotik direnç genlerinin hızlı, hassas ve spesifik saptanması için rutin tanıda alternatif bir tanı yöntemi olarak kullanılabilir.


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Orcid: 0000-0000-0000-0000
Yazar: Zerife ORHAN (Sorumlu Yazar)

Orcid: 0000-0000-0000-0000
Yazar: Arzu KAYIŞ

Orcid: 0000-0000-0000-0000
Yazar: Esra KAYA

Orcid: 0000-0000-0000-0000
Yazar: Murat ARAL

Bibtex @araştırma makalesi { ksutarimdoga452783, journal = {Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi}, issn = {}, eissn = {2619-9149}, address = {Kahramanmaraş Sütçü İmam Üniversitesi}, year = {2018}, volume = {21}, pages = {957 - 965}, doi = {10.18016/ksutarimdoga.vi.452783}, title = {Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus}, key = {cite}, author = {ORHAN, Zerife and ARAL, Murat and KAYA, Esra and KAYIŞ, Arzu} }
APA ORHAN, Z , KAYIŞ, A , KAYA, E , ARAL, M . (2018). Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 21 (6), 957-965. DOI: 10.18016/ksutarimdoga.vi.452783
MLA ORHAN, Z , KAYIŞ, A , KAYA, E , ARAL, M . "Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 (2018): 957-965 <http://dogadergi.ksu.edu.tr/issue/39291/452783>
Chicago ORHAN, Z , KAYIŞ, A , KAYA, E , ARAL, M . "Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 (2018): 957-965
RIS TY - JOUR T1 - Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus AU - Zerife ORHAN , Arzu KAYIŞ , Esra KAYA , Murat ARAL Y1 - 2018 PY - 2018 N1 - doi: 10.18016/ksutarimdoga.vi.452783 DO - 10.18016/ksutarimdoga.vi.452783 T2 - Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi JF - Journal JO - JOR SP - 957 EP - 965 VL - 21 IS - 6 SN - -2619-9149 M3 - doi: 10.18016/ksutarimdoga.vi.452783 UR - http://dx.doi.org/10.18016/ksutarimdoga.vi.452783 Y2 - 2018 ER -
EndNote %0 Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus %A Zerife ORHAN , Arzu KAYIŞ , Esra KAYA , Murat ARAL %T Phenotypic and Genotypic Analysis of Gentamicin, Penicillin, Methicillin, Vancomycin, Linezolid and Tetracycline Resistance in Clinical Isolates of Staphylococcus aureus %D 2018 %J Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi %P -2619-9149 %V 21 %N 6 %R doi: 10.18016/ksutarimdoga.vi.452783 %U 10.18016/ksutarimdoga.vi.452783
ISNAD ORHAN, Zerife , KAYIŞ, Arzu , KAYA, Esra , ARAL, Murat . "Staphylococcus aureus Klinik İzolatlarında Gentamisin, Penisilin, Metisilin, Vankomisin, Linezolid ve Tetrasiklin Direncinin Fenotipik ve Genotipik Olarak Araştırılması". Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi 21 / 6 (Kasım 2018): 957-965. http://dx.doi.org/10.18016/ksutarimdoga.vi.452783