Escherichia coli ve Klebsiella pneumoniea izolatlarında AmpC beta laktamaz sıklığının belirlenmesi ve tespit yöntemlerinin karşılaştırılması / Identifying the frequency of AmpC beta-lactamase Escherichia coli and Klebsiella pneumoniae isolates and comparison of detection methods
Yükleniyor...
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Maltepe Üniversitesi
Erişim Hakkı
CC0 1.0 Universal
info:eu-repo/semantics/openAccess
info:eu-repo/semantics/openAccess
Özet
Bu çalışmanın amacı altın standart olarak PZR yöntemini kullanarak klinik izolatlardan üreyen E.coli ve K. pneumoniae suşlarında plazmid ile kodlanmış AmpC beta-laktamazın araştırılması ve plazmid aracılı AmpC beta-laktamazların saptanması için sefoksitin-Hodge testinin ve kloksasilin inhibisyon testinin etkinliğinin karşılaştırılmasıdır. İki yıl boyunca 3,450 hastanın klinik izolatlarından sefoksitine dirençli 22 E.coli ve 18 K. pneumoniae suşu toplanmıştır. Disk difuzyon testinde sefoksitin dirençli olan suşlara AmpC geni varlığının araştırılması amacı ile MOX,CIT, EBC, FOX, DHA, EBC primerleri kullanılarak multipleks PZR uygulanmıştır. AmpC geni tesbit edilen suşlar Sefoksitin-Hodge testi ve kloksasilin inhibisyon testi ile fenotipik olarak incelenmiştir. 40 izolattan dokuzu, GSBL üretimini teyit eden klavulanik asit tarafından, altısı ise kloksasilin tarafından inhibe edildi, dokuzu sefoksitin-Hodge testi ile pozitif saptandı. Dört izolat, kombinasyon halinde GSBL ve AmpC beta-laktamaz eksprese etti. AmpC betalaktamaz genleri PZR ile incelenmiş, 40 sefoksitine dirençli izolattan 11 tanesinin plasmid aracılı AmpC beta laktamaz sentezlediği görülmüştür (Beş E.coli ve altı K. pneumoniae). 11 izolattan iki CIT, iki EBC, altı FOX, bir izolatta hem FOX ve EBC tipi gen saptanmıştır. İki fenotipik tarama yöntemi kappa analizi ile karşılaştırılmış, Hodge testi plazmid aracılı AmpC laktamazların saptanmasında rehberlerin aksine kloksasilin inhibisyon testinden daha hassas olduğu görülmüştür. Rutin laboratuvarlarda uygulanabilecek daha kolay, ekonomik ve güvenilir bir fenotipik yönteme ihtiyaç olduğu tesbit edilmiştir.
The aim of this study was to look for plasmid encoded AmpC beta-lactamase producing clinical isolates of E.coli and K. pneumoniae using the PCR method as a gold standard and also compare the efficiency of cefoxitin-Hodge test and cloxillin inhibition test for detecting plasmid-mediated AmpC beta-lactamases. Cefoxitin resistant 22 E.coli and 18 K.pneumonia, were collected from 3,450 patient in two years period. Strains which resistant in cefoxitin disk diffusion test investigate for presence of AmpC genes by multiplex PCR using primers MOX CIT, EBC, FOX, DHA, EBC primers. Strains which were positive for AmpC genes investigated by cefoxitin Hodge and cloxacillin disc tests. Among 40 isolates nine were inhibited by clavulanic acid confirming their ESBL production, nine were positive by cefoxitin-Hodge test, six isolates were inhibited by cloxacillin. Four isolates expressed ESBLs and AmpC. Genes encoding for six phylogenic groups acquired AmpC genes were examined by PZR. 11 out of 40 cefoxitin-resistant isolates included AmpC (Five E.coli ve six K. pneumoniae 9). Out of 11 isolates, two produced CIT-type enzymes, two EBC, six FOX and one both FOX and EBC type enzymes. PCR method was most sensitive technique to detect plasmidmediated AmpC lactamases. The two phenotypic screening methods were compared with kappa analysis and the Hodge test was more sensitive than the inhibition test of cloxacillin in the determination of plasmid-mediated AMPC lactamases, unlike the guidelines. It has been determined that there is a need for an easier, economical and reliable phenotypic method that can be applied in routine laboratories.
The aim of this study was to look for plasmid encoded AmpC beta-lactamase producing clinical isolates of E.coli and K. pneumoniae using the PCR method as a gold standard and also compare the efficiency of cefoxitin-Hodge test and cloxillin inhibition test for detecting plasmid-mediated AmpC beta-lactamases. Cefoxitin resistant 22 E.coli and 18 K.pneumonia, were collected from 3,450 patient in two years period. Strains which resistant in cefoxitin disk diffusion test investigate for presence of AmpC genes by multiplex PCR using primers MOX CIT, EBC, FOX, DHA, EBC primers. Strains which were positive for AmpC genes investigated by cefoxitin Hodge and cloxacillin disc tests. Among 40 isolates nine were inhibited by clavulanic acid confirming their ESBL production, nine were positive by cefoxitin-Hodge test, six isolates were inhibited by cloxacillin. Four isolates expressed ESBLs and AmpC. Genes encoding for six phylogenic groups acquired AmpC genes were examined by PZR. 11 out of 40 cefoxitin-resistant isolates included AmpC (Five E.coli ve six K. pneumoniae 9). Out of 11 isolates, two produced CIT-type enzymes, two EBC, six FOX and one both FOX and EBC type enzymes. PCR method was most sensitive technique to detect plasmidmediated AmpC lactamases. The two phenotypic screening methods were compared with kappa analysis and the Hodge test was more sensitive than the inhibition test of cloxacillin in the determination of plasmid-mediated AMPC lactamases, unlike the guidelines. It has been determined that there is a need for an easier, economical and reliable phenotypic method that can be applied in routine laboratories.
Açıklama
Anahtar Kelimeler
kloksasilin AmpC, Escherichia coli, Klebsiella pneumoniae, Plasmid, AmpC, Escherichia coli, Klebsiella pneumoniae, Plasmids
Kaynak
Maltepe Tıp Dergisi
WoS Q Değeri
Scopus Q Değeri
Cilt
11
Sayı
2
Künye
Balıkçı, A. (2019). Escherichia coli ve Klebsiella pneumoniea izolatlarında AmpC beta laktamaz sıklığının belirlenmesi ve tespit yöntemlerinin karşılaştırılması / Identifying the frequency of AmpC beta-lactamase Escherichia coli and Klebsiella pneumoniae isolates and comparison of detection methods. Maltepe Tıp Dergisi. 11(1), s. 23-27.
