Platelet bacterial contamination and the use of a chemiluminescence-linked universal bacterial ribosomal RNA gene probe.
BACKGROUND
Currently, the maximum outdate for platelets is 5 days, because of the increasing chance of bacterial growth over time. Various methods for rapid detection of bacterial contamination of blood components have been described, with mixed results and no general acceptance. A recently described, molecular biologic approach for the detection of bacterial contamination involves a chemiluminescence-linked universal DNA bacterial probe to a highly conserved bacterial region of ribosomal RNA (rRNA).


METHODS
A multicenter trial of a chemiluminescence-linked universal bacterial rRNA probe for the detection of bacterial contamination in platelet concentrates is described. At each of five sites, platelet concentrates (no older than 1 day from date of phlebotomy) were inoculated in triplicate with isolates of four bacterial species (Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus epidermidis, and Staphylococcus aureus) to a final concentration of 10 to 50 colony-forming units (CFUs) per mL and in triplicate to a final concentration of 1000 CFUs per mL. At one site, an additional 6 platelet concentrates were inoculated with sterile saline to serve as controls. Inoculated units were then subjected periodically to quantitative cultures and probe analyses. A total of 126 platelet concentrates were studied over a period of 7 days (120 inoculated with bacteria and 6 with sterile saline).


RESULTS
This assay was, in some cases, able to detect S. aureus bacterial contamination in the range of 100 to 1000 CFUs per mL; the majority of samples (B. cereus, P. aeruginosa, S. aureus, and S. epidermidis) with contamination exceeding 10(4) CFUs per mL; and all samples with contamination of 2.1 x 10(5) CFUs per mL or greater. Increasing the sample size from the recommended 0.4 mL to 1.0 mL resulted in an unacceptable loss of specificity (83.3%).


CONCLUSIONS
The routine use of this assay would be expected to result in a decreased risk of septic platelet transfusion reactions and could lead to a lengthening of the current 5 day storage period for platelets. Further, the pooling of random-donor platelet concentrates before storage instead of immediately before transfusion may be possible if this rRNA probe is employed to detect bacteria in the pool.
Brecher
M E
ME
University of North Carolina at Chapel Hill.
Hogan
J J
JJ
Boothe
G
G
Kerr
A
A
McClannan
L
L
Jacobs
M R
MR
Yomtovian
R
R
Chongokolwatana
V
V
Tegtmeier
G
G
Henderson
S
S
D001409:Bacillus cereus / Q000254:growth & development / Q000302:isolation & purification; D001419:Bacteria / Q000302:isolation & purification*; D001792:Blood Platelets / Q000382:microbiology*; D006801:Humans; D008163:Luminescent Measurements; D017713:Platelet Transfusion; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000302:isolation & purification; D015347:RNA Probes; D012329:RNA, Bacterial / Q000032:analysis* / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D013211:Staphylococcus aureus / Q000254:growth & development / Q000302:isolation & purification; D013212:Staphylococcus epidermidis / Q000254:growth & development / Q000302:isolation & purification
D015347:RNA Probes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal