The use of a chemiluminescence‐linked universal bacterial ribosomal RNA gene probe and blood gas analysis for the rapid detection of bacterial contamination in white cell‐reduced and nonreduced platelets

Abstract

Because of the rising incidence of bacterial growth and septic platelet transfusions in aging units, platelet storage is currently limited in the United States to 5 days. This approved shelf life of platelets might be altered if methods were devised to rapidly detect infected units and/or to decrease the incidence of bacterially contaminated platelets. An investigation was conducted on the effect of a prototype blood collection system with an in-line filter for the production of white cell-reduced platelet-rich plasma on the growth of bacteria in platelets prepared from whole blood that had been inoculated with Staphylococcus epidermidis. Additional studies were conducted with a chemiluminescence-linked ribosomal RNA (rRNA) gene probe and with blood gas analysis to identify possible methods for the rapid detection of bacterial contamination. All units were followed for 9 days of storage. The filtration of the platelet-rich plasma resulted in an approximate 2 log10 reduction in white cells with an average loss of 6.7 percent of platelets. Filtration did not appear to alter bacterial growth. In all platelet units that supported growth, pO2 dropped to negligible values and pCO2 rose relative to culture-negative units. The changes were most sensitive and specific beyond 5 days of storage. The universal bacterial rRNA probe assay was able to detect S. epidermidis in concentrations as low as 1 x 10(3) colony-forming units per mL in some cases and reliably detected all units contaminated at a concentration of 1 x 10(4) colony-forming units per mL.(ABSTRACT TRUNCATED AT 250 WORDS)

Transfusion-related bacterial sepsis

“…As expected, S. epidermidis, S. aureus, Bacillus cereus, E. coli, and other organisms that produce acid as a by-product of glycolysis significantly lowered extracellular pH by day 3 of storage. Similar results with S. epidermidis have been recently obtained by Brecher and colleagues, who also demonstrated that differences between the control and contaminated platelets were evident when titers reached a level of 3 x 106 CFU/ml (18).…”

“…Sherburne and colleagues failed to observe differences between the titers of S. aureus in control and leukodepleted (using a Pall PL-50 filter [Pall Biomedical, Glen Cove, N.Y.]) platelet concentrates (prepared from inoculated platelet-rich plasma that was subsequently leukodepleted) (124). Brecher and colleagues and Wenz and coworkers also failed to observe reduction in bacterial levels following leukodepletion of contaminated platelet concentrates (18,150).…”

“…Several investigators have described a novel application of a chemiluminescence-linked rRNA gene probe system to detect bacterial contamination in platelet concentrates (18,27,60,74 (46,136,143).…”

Bacterial Contamination of Blood Components

“…Because of the high sensitivity and specificity and the rapidity of obtaining results, molecular biological approaches are theoretically appealing for the detection of contaminating organisms in blood components. One such approach is the use of a nonamplified chemiluminescence-linked universal bacterial rRNA probe (5,9). This technique was shown to detect four bacterial species in the range of 10 4 to 10 5 CFU/ml.…”

Rapid Diagnosis of Bacterial Infections Using Technologies Based on Nucleic Acid Detection

“…Genes highly conserved through bacterial evolution are always involved in essential functions of the cell and are constantly challenged by a strong selective pressure. These highly conserved genes offer great potential for the universal detection of bacteria or for detection at the genus or family levels (14)(15)(16)(17)(18)(19). Furthermore, highly conserved nucleotide sequences suitable for universal detection of bacteria or for bacterial detection at the genus or family levels may be interspersed by more variable sequences that may be exploited for species-specific detection of bacteria (15)(16)(17).…”