Background: The implementation of diversion pouches is to minimise the risk of bacterial contamination as the initial blood flow is prevented from entering primary bag collections as it is diverted into a pouch. This study was carried out to determine the prevalence of bacterial contamination in the diversion pouches used  during blood collections in the Transfusion Department of Hospital Seberang Jaya, Penang, Malaysia. Methods: BD Bactec™ Fx instrument detection system was performed on 702 samples of 20 mL of  diverting blood in diversion pouch. The  inocullum  volume was 10 mL for both aerobic and anaerobic bottles cultures and incubated for 5 days in  the BD Bactec™ Fx instrument. Positive sample was flagged by BD Bactec™ Fx instrument and  subculture  to identify the species of organism. Results: The results  showed that of 702 samples, 12 (1.7%) were contaminated. The bacterial species identified were coagulase negative Staphylococcus, Staphylococcus aureus and Gram positive Bacilli. Conclusion: The results  strongly suggest that the usage of diversion pouch is of significant importance in reducing bacterial contamination during blood collection.