Performance and suitability of polymerase chain reaction for early detection of bacteria in platelet concentrates

Transfusion. 2011 Sep;51(9):2006-11. doi: 10.1111/j.1537-2995.2011.03090.x. Epub 2011 Mar 10.

Abstract

Background: In this study the applicability of a 16S rRNA real-time reverse transcriptase polymerase chain reaction (RT-PCR) and a Staphylococcus genus-specific PCR for screening of bacterial contamination in platelet concentrates (PCs) was determined.

Study design and methods: A total of 336 sample bags, from PCs that were routinely tested in the BacT/ALERT (bioMérieux), were collected and frozen until testing by the PCR assays. Based on the BacT/ALERT results, 107 PCs were positive and 229 were negative for bacterial growth.

Results: The analytical sensitivity of the 16S rRNA real-time RT-PCR ranged from 5 to 40 colony-forming units (CFUs)/mL. The PCR detected five positive samples, four of which were also positive in the BacT/ALERT. The sensitivity of the test was 3.8%, and the specificity was 99.5%. The analytical sensitivity of the Staphylococcus genus-specific PCR ranged from 5 to 15 CFUs/mL. Thirty-nine units that were BacT/ALERT positive for staphylococci were tested with this PCR. Six samples were positive with the PCR, five of which were also BacT/ALERT positive. The sensitivity of the Staphylococcus genus-specific PCR was 12.8%, and the specificity was 98.8%.

Conclusion: Despite the rapid availability of results compared to the BacT/ALERT, the analytical sensitivity of a generic or specific PCR assay is not high enough to be an alternative for the BacT/ALERT when PCs are screened on the day of production.

MeSH terms

  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Blood Platelets / microbiology*
  • Humans
  • RNA, Ribosomal, 16S / genetics
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • RNA, Ribosomal, 16S