Akut ishalli beş yaş altı çocuklarda diyarejenik escherichia coli izolatlarının sıklığı
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Akut ishalli beş yaş altı çocuklarda, izole edilen Escherichia coli tiplerinin tespiti, hastalığın tanısı ve tedavisi açısından önemlidir. Türkiye'de çocuklarda diyarejenik E. coli patotiplerinin sıklığı hakkında çalışma ve veri bulunmamaktadır. Bu çalışmanın amacı Konya/Türkiye'de, beş yaş altı çocuklarda 2019 yılında, akut ishale yol açan diyarejenik E. coli patotiplerinin [enteropatojenik E. coli (EPEC), enterotoksijenik E. coli (ETEC), enteroinvazif E. coli (EIEC), Shiga toksin üreten E. coli, enterohemorajik E. coli (EHEC), enteroagregatif E. coli (EAEC) ve diffüz aderan E. coli (DAEC)] sıklığının araştırılmasıdır. Gereç ve Yöntem Selçuk Üniversitesi Tıp Fakültesi Hastanesi, Tıbbi Mikrobiyoloji Laboratuvarı'na 1 Ocak-31 Aralık 2019 tarihleri arasında gönderilen akut ishalli beş yaş altı çocuklardan alınan 3064 dışkı örneği tarandı. Salmonella, Shigella, Campylobacter üremeyen, rotavirus ve adenovirus pozitifliği saptanmayan ve parazit görülmeyen 312 çocuğun dışkısında baskın üreyen E.coli izolatları çalışmaya alındı. E. coli konvansiyonel yöntemler ve Vitek 2 compact otomatize sistemi ile tanımlandı. DNA izolasyonu, ticari bir kitle yapıldı. eae, pEAF, daaC, aatA, elt, est, ipaH, stx1 ve stx2 primerleri gerçek zamanlı multipleks PZR yöntemi ile çalışıldı. Bir izolatta eae genin saptanması aEPEC, eae ve pEAF genlerinin saptanması tEPEC, stx1 ve/veya stx2 genlerinin saptanması STEC, eae, stx1 ve/veya stx2 genlerinin saptanması EHEC, elt ve/veya est genlerinin saptanması ETEC, ipaH genin saptanması EIEC, aatA genin saptanması EAEC ve daaC geninin saptanması DAEC olarak tanımlandı. Gerçek zamanlı multipleks PZR, monopleks PZR ve jel elektroforezi ile doğrulandı. Bulgular: Toplam 312 E. coli izolatının 56'sında tek, 254'ünde miks patotip saptanırken iki izolat tiplendirilemedi. 310 örnekte 281 DAEC, 187 EPEC, 159 EAEC, 43 EIEC, 31 EHEC, dört ETEC ve iki STEC saptandı. En sık tespit edilen tekli patotipler DAEC (47) ve EAEC (7) iken miks patotipler DAEC + EPEC + EAEC (108), DAEC + EPEC (58) ve DAEC + EIEC (24) olarak belirlendi. Sonuç: Konya/Türkiye'de beş yaş altı akut ishalli çocuklarda DAEC'in baskın patotip olduğu ve miksenfeksiyonlar da ise en sık DAEC+EPEC+EAEC birlikteliği saptanmıştır. Bu çalışma Türkiye'de tüm diyarejenik E.coli patotiplerinin araştırıldığı ilk çalışmadır. Çalışmamızın en önemli kısıtlılığı tek merkezli olmasıdır. Türkiye'de hangi patotiplerin yaygın olduğunun belirlenmesi için çok merkezli çalışmalara ihtiyaç vardır.
The detection of diarrheagenic Escherichia coli isolates from normal intestinal microbiota is important for the diagnosis and treatment of acute diarrhea. There is no research and data about the frequency of diarrheagenic E. coli pathotypes in Turkey. This study aims to detect the frequency of diarrheagenic E. coli pathotypes [enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) Shiga toxin-producing E. coli, enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC) and diffuse adherence E. coli (DAEC)] in children with acute diarrhea under five years in Konya/Turkey. Material and Method: 3064 stool samples taken from children with acute diarrhea under the age of five years sent to the Medical Microbiology Laboratory of the Medical Faculty Hospital, Selcuk University between 1 January and 31 December 2019 were scanned. E.coli isolates were included in the study that predominantly growing in the feces of 312 children whose Salmonella, Shigella, Campylobacter were not grow, rotavirus and adenovirus positivity was not detected and no parasites were observed. E. coli was defined by conventional methods and Vitek 2 compact automated system. DNA isolation was performed with a commercial kit. The primers of eae, pEAF, daaC, aatA, elt, est, ipaH, stx1 and stx2 genes were studied by real-time multiplex PCR method. Detection of eae plus pEAF genes was accepted typical EPEC (tEPEC), only eae atypical EPEC (aEPEC), stx1 and/or stx2 genes STEC, eae plus stx1 and/or stx2 genes EHEC, elt and/or est genes ETEC, ipaH gene EIEC, aatA gene EAEC and daaC gene DAEC. Results were confirmed by monoplex PCR and gel electrophoresis. Results: Of the 312 E.coli isolates, 56 had single pathotype, 254 had mixed pathotypes and two isolates could not be typed. 281 DAEC, 187 EPEC, 159 EAEC, 43 EIEC, 31 EHEC, four ETEC and two STEC were detected in 312 samples. The most frequently detected single pathotypes were DAEC (47) and EAEC (7), while mixed pathotypes were DAEC + EPEC + EAEC (108), DAEC + EPEC (58) and DAEC + EIEC (24) respectively. Conclusion: DAEC was single and also DAEC plus tEPEC/aEPEC plus EAEC were mixed predominant pathotypes in the diarrheagenic E. coli isolates in children with acute diarrhea under five years in Konya/Turkey. This is the first study to investigate all diarrheagenic E. coli pathotypes in Turkey. The most important limitation of our study is that single-centered. Multicentre studies are needed to determine which pathotypes are common in Turkey.
The detection of diarrheagenic Escherichia coli isolates from normal intestinal microbiota is important for the diagnosis and treatment of acute diarrhea. There is no research and data about the frequency of diarrheagenic E. coli pathotypes in Turkey. This study aims to detect the frequency of diarrheagenic E. coli pathotypes [enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) Shiga toxin-producing E. coli, enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC) and diffuse adherence E. coli (DAEC)] in children with acute diarrhea under five years in Konya/Turkey. Material and Method: 3064 stool samples taken from children with acute diarrhea under the age of five years sent to the Medical Microbiology Laboratory of the Medical Faculty Hospital, Selcuk University between 1 January and 31 December 2019 were scanned. E.coli isolates were included in the study that predominantly growing in the feces of 312 children whose Salmonella, Shigella, Campylobacter were not grow, rotavirus and adenovirus positivity was not detected and no parasites were observed. E. coli was defined by conventional methods and Vitek 2 compact automated system. DNA isolation was performed with a commercial kit. The primers of eae, pEAF, daaC, aatA, elt, est, ipaH, stx1 and stx2 genes were studied by real-time multiplex PCR method. Detection of eae plus pEAF genes was accepted typical EPEC (tEPEC), only eae atypical EPEC (aEPEC), stx1 and/or stx2 genes STEC, eae plus stx1 and/or stx2 genes EHEC, elt and/or est genes ETEC, ipaH gene EIEC, aatA gene EAEC and daaC gene DAEC. Results were confirmed by monoplex PCR and gel electrophoresis. Results: Of the 312 E.coli isolates, 56 had single pathotype, 254 had mixed pathotypes and two isolates could not be typed. 281 DAEC, 187 EPEC, 159 EAEC, 43 EIEC, 31 EHEC, four ETEC and two STEC were detected in 312 samples. The most frequently detected single pathotypes were DAEC (47) and EAEC (7), while mixed pathotypes were DAEC + EPEC + EAEC (108), DAEC + EPEC (58) and DAEC + EIEC (24) respectively. Conclusion: DAEC was single and also DAEC plus tEPEC/aEPEC plus EAEC were mixed predominant pathotypes in the diarrheagenic E. coli isolates in children with acute diarrhea under five years in Konya/Turkey. This is the first study to investigate all diarrheagenic E. coli pathotypes in Turkey. The most important limitation of our study is that single-centered. Multicentre studies are needed to determine which pathotypes are common in Turkey.
Açıklama
Anahtar Kelimeler
Akut İshal, Diyarejenik Escherichia Coli, Diarrheagenic Escherichia Coli, Acute Diarrhea, Multiplex Polymerase Chain Reaction
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Koç, A. (2020). Akut ishalli beş yaş altı çocuklarda diyarejenik escherichia coli izolatlarının sıklığı. (Yüksek Lisans Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
