Sixty-eight patients with AP and 157 healthy controls were recruited from the archives of the Department of Gastroenterology, Ege University Medical School, İzmir, Turkey. Informed written consent was obtained from each patient and the study protocol conforms to the ethical guidelines of the declaration of Helsinki. Etiology, gender, age, clinical presentation, clinical course, genotype frequency of the ACE I/D polymorphism and their association were compared.

 

Blood specimens from all participants were obtained with a standard venipuncture technique that used ethylenediamine tetra-acetic acid-containing tubes. DNA was isolated from peripheral blood by a standard phenol/chloroform extraction method [12].

 

For the ACE polymorphism, we used polymerase chain reaction (PCR) methodology [19], with the upstream primer 5’-CTG GAG ACC ACT CCC ATC CTT TCT-3’ and the downstream primer 5’-GAT GTG GCC ATC ACA TTC GTC AGA T-3’. Amplification was performed for 35 cycles with denaturation, extension and annealing temperatures of 94.8°C, 60.8°C and 72.8°C, respectively. The resulting PCR products were separated on 2% agarose gel with ethidium bromide staining and were visualized under ultraviolet light. Homozygotes for the deletion or insertion genotypes were described as DD and II, respectively, and the heterozygous genotype as ID.