Technical Abstract: The effect of pooling five or 11 orophyarngeal (O/P) swabbings on detecting avian influenza virus (AIV) by real-time reverse transcription-polymerase chain reaction (RRT-PCR) was evaluated. The model used for the evaluation was designed to minimize viral load and thus assess the effect of the pooling on detection. Two-week-old broiler chickens were inoculated via the intranasal route with the low pathogenicity (LP) chicken/Maryland/Minh Ma/04 H7N2 strain or remained uninoculated. On day 2, 3, 4, 5, 7, 9, 11, and 14 post-inoculation (PI), O/P swabbings were collected from individual infected birds and pooled with either four or ten O/P swabs from uninfected broilers to produce ten replicate pools of five and 11 swabbings, respectively. AIV was readily detected (80-100%) by RRT-PCR in the pools of five and pools of eleven swabbings from day 2 through day 5 PI. Detection in pools of both types decreased to similar levels on Day 7 (50% for the pools of 5 and 40% for the pools of 11). AIV was not detected on day 9, 11, and 14 PI. On a given sample day PI, mean cycle threshold (Ct) values were consistently lower (higher genome levels) in the pools of five compared to the pools of 11. These differences were statistically significant only on day 3 and 5 PI, yet Ct values were both types of pools were clearly interpretable as AIV positive.