Çeşitli escherichia coli suşlarının pulsed field jel elektroforez yöntemi ile tiplendirilmesi, plazmit profilleri ve antibiyotik duyarlılıklarının araştırılması
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Tarih
2012-04-11
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Selçuk Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada; kuyu suları (43), çeşitli gıdalar (40) ve klinik örneklerden (47) izole ve identifiye edilen toplam 130 Escherichia coli suşu; genetik çeşitlilik, bazı antibiyotiklere duyarlılık ve plazmit profilleri yönünden araştırıldı. E. coli izolatlarının genetik ilişkilerini ve çeşitliliklerini tanımlamak için Pulsed-field jel elektroforezi (PFGE) metodu kullanıldı. PFGE sonuçları % 80 benzerlik oranına göre değerlendirildiğinde; E. coli suşları, 16 alt tiple temsil edilen 101 farklı pulsotipe ayrıldı. Analiz edilen 43 su izolatı arasında 30 farklı pulsotip ve altı alt tip görülürken; 40 gıda suşu 32 farklı pulsotipe ve altı alt tipe, 47 klinik izolat ise 39 farklı pulsotip ve dört alt tipe ayrıldı. Restriksiyon profilleri değerlendirildiğinde tüm suşlar arasında yüksek oranda genetik çeşitlilik görüldü. Genotipik analizler, farklı gruplara ait olan suşların köken olarak aynı olmadığını ve ilişkisiz izolat olarak belirlendiğini gösterdi. Antibiyotik duyarlılık testleri 15 farklı antibiyotiğe karşı, disk difüzyon yöntemi ile yapıldı. Su izolatlarının en fazla direnç gösterdiği antibiyotiğin ampisilin (% 97.67) olduğu; bunu % 9.3 ile tikarsilin/klavulanik asit, % 6.97 ile sefuroksim ve seftazidim'in takip ettiği görüldü. Gıda izolatlarının direnç oranlarının ampisilin'e karşı % 77.5, tikarsilin/klavulanik asit'e karşı % 30, tetrasiklin'e karşı % 22.5 ve sefalotin'e karşı % 14.5 olduğu belirlendi. Klinik izolatlarda direnç oranları; ampisilin'e karşı % 100, tikarsilin/klavulanik asit ve sefalotin'e karşı % 89.36, sefuroksim'e karşı % 87.24, tetrasiklin ve sefotaksim'e karşı % 80.85 olarak belirlendi. Suşların plazmit izolasyonları, alkali lizis yöntemi kullanılarak yapıldı. 43 su izolatının 19'u ( % 44.18), 78.2-2.6 kb aralığında değişen boyutta 19 farklı plazmit bandı ve dört farklı plazmit profili gösterdi. 40 gıda izolatının 18'i (% 45), 64-1 kb arasında değişen boyutta 31 farklı plazmit bandı ve yedi farklı plazmit profili gösterirken; 47 klinik izolatın 27'si (% 57.44), 77.1-1.6 kb aralığında değişen boyutta 57 farklı plazmit bandı ve 22 farklı plazmit profili gösterdi. Üç grup için aynı boyutta plazmit belirlenmedi. İzole edilen E. coli suşlarında plazmitlerin rastgele dağılım gösterdiği ve antibiyotik direnç profilleri ile plazmitler arasında önemli bir ilişki kurulamadığı sonucuna varıldı.
In this study, a total of 130 Escherichia coli strains isolated and identified from well waters (43), various foods (40) and clinical samples (47) were investigated in terms of their genetic diversity, susceptibility against some antibiotics and plasmid profiles. Pulsed-field gel electrophoresis method (PFGE) was used to characterize the genetic relationship and diversity of E. coli isolates. When the results of PFGE were evaluated according to similarity level of 80 %, the E. coli strains were seperated into 101 distinct groups representing 16 subtypes. While 30 different pulsotype and six different subtypes were revealed among 43 water isolates analyzed; 40 food isolates and 47 clinical isolates were divided into 32 and 39 different pulsotype, six and four subtypes, respectively. When the restriction profiles were evaluated, a great genetic diversity was seen among all strains. Genotypic analysis indicated that strains which belong to different groups were not identical and determined as unrelated strains. Antibiotic susceptibility tests were performed by using disc diffusion methods against 15 different antibiotics. Ampicillin was the most common antibiotic that water isolates resistant to (97.67 %), followed by ticarcillin/clavulanic acid (9.3 %), cefuroxime and ceftazidime (6.97 %). Resistance rates of food isolates were determined against ampicillin as 77.5 %, against ticarcillin/clavulanic acid 30 %, against tetracycline 22.5 % and against cephalothin 14.5 %. Resistance rates of clinical isolates were determined as 100 % against ampicillin, followed by 89.36 % against ticarcillin/clavulanic acid and cephalothin, 87.24 % against cefuroxime, 80.85 % against tetracycline and cefotaxime. Plasmid isolations of the strains were performed by using alkaline lysis method. 19 of 43 water isolates (44.18 %) exhibited 19 different plasmid bands in the range of 78.2 -2.6 kb and four different plasmid profiles. Although 18 of 40 food isolates (45 %) showed 31 different plasmid bands in the range of 64-1 kb and seven different plasmid profiles, 27 of 47 clinical isolates (57.44 %) revealed 57 different plasmid bands in the range of 77.1 kb-1.6 kb and 22 different plasmid profiles. Identical plasmid sizes were not detected for all three groups. It was concluded that the plasmids were distributed at random in the isolated E. coli strains and no remarkable relationship between antibiotic resistance patterns and plasmids could be established.
In this study, a total of 130 Escherichia coli strains isolated and identified from well waters (43), various foods (40) and clinical samples (47) were investigated in terms of their genetic diversity, susceptibility against some antibiotics and plasmid profiles. Pulsed-field gel electrophoresis method (PFGE) was used to characterize the genetic relationship and diversity of E. coli isolates. When the results of PFGE were evaluated according to similarity level of 80 %, the E. coli strains were seperated into 101 distinct groups representing 16 subtypes. While 30 different pulsotype and six different subtypes were revealed among 43 water isolates analyzed; 40 food isolates and 47 clinical isolates were divided into 32 and 39 different pulsotype, six and four subtypes, respectively. When the restriction profiles were evaluated, a great genetic diversity was seen among all strains. Genotypic analysis indicated that strains which belong to different groups were not identical and determined as unrelated strains. Antibiotic susceptibility tests were performed by using disc diffusion methods against 15 different antibiotics. Ampicillin was the most common antibiotic that water isolates resistant to (97.67 %), followed by ticarcillin/clavulanic acid (9.3 %), cefuroxime and ceftazidime (6.97 %). Resistance rates of food isolates were determined against ampicillin as 77.5 %, against ticarcillin/clavulanic acid 30 %, against tetracycline 22.5 % and against cephalothin 14.5 %. Resistance rates of clinical isolates were determined as 100 % against ampicillin, followed by 89.36 % against ticarcillin/clavulanic acid and cephalothin, 87.24 % against cefuroxime, 80.85 % against tetracycline and cefotaxime. Plasmid isolations of the strains were performed by using alkaline lysis method. 19 of 43 water isolates (44.18 %) exhibited 19 different plasmid bands in the range of 78.2 -2.6 kb and four different plasmid profiles. Although 18 of 40 food isolates (45 %) showed 31 different plasmid bands in the range of 64-1 kb and seven different plasmid profiles, 27 of 47 clinical isolates (57.44 %) revealed 57 different plasmid bands in the range of 77.1 kb-1.6 kb and 22 different plasmid profiles. Identical plasmid sizes were not detected for all three groups. It was concluded that the plasmids were distributed at random in the isolated E. coli strains and no remarkable relationship between antibiotic resistance patterns and plasmids could be established.
Açıklama
Anahtar Kelimeler
Antibiyotik duyarlılık testi, Escherichia coli, Plazmit profili, Pulsedfield jel elektroforezi, Antibiotic susceptibility test, Plasmid profile, Pulsed-field gel electrophoresis
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Künye
Uysal, A. (2012). Çeşitli escherichia coli suşlarının pulsed field jel elektroforez yöntemi ile tiplendirilmesi, plazmit profilleri ve antibiyotik duyarlılıklarının araştırılması. Selçuk Üniversitesi, Yayımlanmış doktora tezi, Konya.
