2016 ACVIM Forum Research Abstract Program

A minimum of seven dogs in each group (normal versus CHF) were needed to demonstrate the statistical power with a probability of 0.9 and type one error of P < 0.05 based on previous publications. Blood was collected in EDTA tubes from 9 normal geriatric dogs (free of heart murmur and absent history of CHF) and 8 dogs with CHF secondary to MMVD (confirmed by clinical signs, chest x-ray and echocardiography). Plasma samples were subjected to isolation of miRNAs with a miRNeasy Plasma Kit (Qiagen) and subsequently reverse-transcription reaction with a miScript II RT Kit (Qiagen). Validated canine primers of 12 candidate miRNAs and miR-39 (spike-in control) were employed to quantify relative expression levels of each miRNA between normal and CHF dogs via real time PCR (miScript SYBR Green PCR Kit, Qiagen).