Environmental swabs from pullet houses were tested for the presence of Salmonella Enteritidis (SE) using the RapidChek SE immunoassay test. Of the 1162 samples tested in 2011, 20 samples were positive, but only two samples were confirmed positive for SE by culture. Seventeen positive samples were from pullet houses that had been vaccinated with SE bacterin 2 to 3 days prior to submission to the lab. This study investigated the detection of SE bacterin using the RapidChek SE test system. Swabs were inoculated with different amounts of a commercial SE bacterin. As little as 0.25 ml of the SE bacterin was sufficient to cause positive results with the RapidChek SE test. This finding emphasizes the need to perform bacterial isolation and identification when a positive result occurs using an immunoassay test.