Gökhan İNAT, Ertan Emek ONUK, Arzu FINDIK, Alper ÇİFTCİ, İnci Ülkü ÇELİKEL

Sığır karkas orijinli Escherichia coli O157 izolatlarının rastgele çoğaltılmış polimorfik DNA-polimeraz zincir reaksiyonu ile genotiplendirilmesi

Bu çalışma, sığır karkas orijinli 32 adet Escherichia coli O157 izolatının iki farklı random primerin (M13 ve ERIC2) kullanıldığı rastgele çoğaltılmış polimorfik DNA-polimeraz zincir reaksiyonu (RAPD-PCR) metodu ile genotiplendirilmesi ve farklı primer bağlanma ısılarının (32, 36, 38 ve 40°C) RAPD bant paternleri üzerine etkisinin değerlendirilmesi amacıyla yapılmıştır. İzolatlar M13 primeri ile 7, ERIC2 primeri ile 5 farklı genotip içerisinde dağıldı. Çalışmada kullanılan primer bağlanma ısılarının RAPD bant paternlerinin çeşitliliğini dikkate değer bir şekilde etkilemediği görüldü. Sonuç olarak E. coli O157 izolatları arasındaki çeşitliliklerin belirlenmesinde M13 primerinin daha uygun olduğu ve epidemiyolojik çalışmalarda kullanılabilir olduğu belirlendi.

Anahtar Kelimeler:

Alper ÇİFTCİ, İnci Ülkü ÇELİKEL, Arzu FINDIK, Ertan Emek ONUK, Gökhan İNAT, Escherichia coli O157, genotiplendirme, primer bağlanma ısısı, RAPD-PCR., 2010, Etlik Veteriner Mikrobiyoloji Dergisi

Genotyping of beef carcass isolates of Escherichia coli O157 by random amplified polymorphic DNA-polymerase chain reaction

This study was performed to genotype 32 E. coli O157 isolates originating from bovine carcasses by random amplified polimorphic DNA-polymerase chain reaction (RAPD-PCR) methods using two different random primers (M13 and ERIC2) and to evaluate the effect of the different (32, 36, 38 and 40°C) annealing temperatures on RAPD band patterns. Isolates were distributed into 7 and 5 different genotypes using M13 and ERIC2 primers, respectively. It was observed that different annealing temperatures used in this study had not considerable effect on RAPD band variability. In conclusion, it was determined that M13 primer was more convenient to detect genetic variability among E. coli O157 isolates and was useful in epidemiological studies.

Keywords:

Escherichia coli O157, genotyping, primer annealing temperature, RAPD-PCR.,

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