Microbial screening of UC blood units by an automated culture system: Effect of delayed testing on bacterial detection

Background. Microbial screening is a mandatory test for banked UC blood (UCB) to comply with the code of good manufacturing practice (GMP). Cord blood banks (CBBs) are not always closely located to a GMP-licensed microbiology laboratory, resulting in time delays for transport of specimens prior to microbiological testing. This study investigated the influence of ≥ 24 h delays in initiating automated microbial screening on the detection of bacteria in UCB, by analysis of specimens deliberately spiked with bacteria and the recovery of bacteria from cryopreserved spiked-UCB. Materials and methods. UCB was processed according to standard CBB procedures and spiked with Staphylococcus epidermidis or Escherichia coli [2-2000 colony forming units (CFU)/mL]. Spiked-UCB (0.5 mL) was (1) held at room temperature (RT) and inoculated into pediatric BacT/Alert bottles (bioMérieux) at Days 1, 4 and 7 (delayed inoculation); and (2) inoculated directly (Day 0) into replicate BacT/Alert bottles and held at RT for 1, 4 or 7 days before loading onto the BacT/ALERT system (delayed loading). Spiked-UCB samples were cryopreserved. Bacterial counts were quantitated on horse blood agar plates. Results. Bacterial growth in UCB spiked with a single bacterium was capable of detection by the BacT/ALERT system. S. epidermidis grew in all conditions of delayed testing (ie. delayed inoculation and delayed loading). E. coli failed to grow under conditions of delayed inoculation but grew at all time points of delayed loading. S. epidermidis and E. coli were recovered from cryopreserved spiked-UCB. Discussion. Inoculation of culture bottles as soon as possible after sample preparation is preferable. Bacteria can maintain viability in BacT/ALERT bottles inoculated and held at RT for up to 7 days prior to automated culture testing. Bacteria can be successfully recovered from cryopreserved UCB.