Consecutive BacT/Alert blood cultures which were instrument negative following a 7-day incubation were subcultured. Eighteen (0.2%) of 11,476 bottles had growth on subculture. Eleven of these eighteen isolates were considered contaminants on the basis of the identity of the organism and lack of other positive blood cultures from the same patient. In addition, analysis of time to instrument detection for approximately 2,900 positive blood cultures indicates that 5 or 6 days of incubation is sufficient for the routine detection of clinically significant organisms from BacT/Alert blood cultures. These data indicate that subculture of 5- to 7-day instrument-negative BacT/Alert blood culture bottles is not necessary.