Evaluation of two detection methods of microorganisms in platelet concentrates

Evaluation of two detection methods of microorganisms in platelet concentrates

Author Albertoni, G. Google Scholar
Andrade, S. S. Autor UNIFESP Google Scholar
Araujo, P. R. B. Google Scholar
Carvalho, F. O. Google Scholar
Girao, M. J. B. C. Autor UNIFESP Google Scholar
Barreto, J. A. Google Scholar
Institution Colsan Assoc Beneficente Coleta Sangue
Universidade Federal de São Paulo (UNIFESP)
Abstract Background: the performance of a bacterial 16S ribosomal DNA real-time polymerase chain reaction (PCR) assay was evaluated and validated with an automated culture system to determine its use for screening of platelet concentrates (PCs).Study Design and Methods: PCs were spiked with suspensions of Escherichia coli, Serratia marcescens, Staphylococcus epidermidis and St. aureus at 1, 10, and 100 colony-forming units (CFUs) mL and stored for 5 days. DNA amplification was performed using real-time PCR. the BacT/ALERT was used as a reference method and samples were inoculated into an aerobic culture bottle; for the PCR assay, aliquots were drawn from all (spiked) PCs on days 0 to 5 of storage.Results: Real-time PCR detected only the gram-positive bacteria in PCs spiked with low bacterial titres (1 CFU mL) after 48 h; however, it was able to detect all positive samples in PCs spiked with 10 CFU mL of either gram-positive or gram-negative bacteria after 48 h. in addition, real-time PCR detected all positive samples in PCs spiked with high gram-positive bacterial titres (100 CFU mL) after 24 h. On the other hand, the BacT/ALERT system showed positive results in all samples within 24 h.Conclusion: the BacT/ALERT method is more sensitive and should continue to be the gold standard for identifying bacterial contaminations in blood samples. the real-time PCR approach can be used for the screening of PCs for microbial detection before they are released from blood centres or shortly before they are used in blood transfusion, and thus allow an extended shelf life of the platelets.
Keywords BacT/ALERT
bacterial contamination
blood components
molecular testing
platelet concentrates
real-time PCR
Language English
Sponsor Associacao Beneficente de Coleta de Sangue (Colsan)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Financiadora de Estudos e Projetos (FINEP)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Date 2011-12-01
Published in Transfusion Medicine. Hoboken: Wiley-Blackwell, v. 21, n. 6, p. 408-416, 2011.
ISSN 0958-7578 (Sherpa/Romeo, impact factor)
Publisher Wiley-Blackwell
Extent 408-416
Origin http://dx.doi.org/10.1111/j.1365-3148.2011.01105.x
Access rights Closed access
Type Article
Web of Science ID WOS:000297752300010
URI http://repositorio.unifesp.br/handle/11600/34315

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