Detection of bacteria in platelet concentrates: comparison of broad‐range real‐time 16S rDNA polymerase chain reaction and automated culturing

Abstract

Compared to culture in the BacT/ALERT system, the PCR assay had a sensitivity of 100 percent and a specificity of 100 percent. This real-time PCR assay has a much shorter turnaround time of 4 hours, which offers the possibility to test and obtain results on PCs before release or the day they are transfused. This would permit the withdrawal of contaminated PCs before transfusion.

High-Volume Extraction of Nucleic Acids by Magnetic Bead Technology for Ultrasensitive Detection of Bacteria in Blood Components

“…These techniques require centrifugation or vacuum steps and often have lengthy processing times, as well as volume limitations. Covering magnetic beads with nucleic acid-binding matrices is the basis of a different methodologic approach (26 ). Binding to magnetic beads allows the generic (not sequence-specific) isolation of nucleic acids.…”

Transfusion-Associated Bacterial Sepsis

“…Other genetic methods use probes directed at a precisely defined bacterial DNA fragment, mainly 16SrDNA. They enable many different bacteria to be detected in blood components (63). The presence of marked probes is detected with chemiluminescence or electroluminescence.…”

Broad‐Range PCR for Detection and Identification of Bacteria

“…Bacterial broad-range PCR has also attracted interest in transfusion medicine, in order to assess blood products for bacterial contamination (35,36,97,126). Platelet concentrates are of particular concern, because they require storage and agitation at room temperature, thus making them prone to bacterial growth from very small inocula.…”

“…may require 4 to 5 days, so that cultures may become positive at a time when platelets have been transfused. Several broad-range real-time PCR assays have been designed and evaluated for that purpose (35,36,97,126). One study evaluated 2,146 platelet concentrate samples in parallel with culture and PCR (97).…”

“…Several broad-range real-time PCR assays have been designed and evaluated for that purpose (35,36,97,126). One study evaluated 2,146 platelet concentrate samples in parallel with culture and PCR (97). The test had a turnaround time of 4 h and showed concordant results between the two methods, thus showing the feasibility of PCR testing for platelet concentrates.…”