| Author | Alexandrino, Fabiana | |
| Author | Malgarin, Juliane Soldi | |
| Author | Krieger, Marco Aurélio | |
| Author | Morello, Luis Gustavo | |
| Access date | 2020-09-23T16:40:04Z | |
| Available date | 2020-09-23T16:40:04Z | |
| Document date | 2020 | |
| Citation | ALEXANDRINO, F. et al. Optimized broad-range real-time PCR-based method for bacterial screening of platelet concentrates. Brazilian Journal of Biolog. p. 1-9, 2020. | pt_BR |
| ISSN | 1678-4375 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/43572 | |
| Language | por | pt_BR |
| Publisher | Instituto Internacional de Ecologia | pt_BR |
| Rights | open access | pt_BR |
| Title | Optimized broad-range real-time PCR-based method for bacterial screening of platelet concentrates | pt_BR |
| Type | Article | pt_BR |
| Abstract | Bacterial contamination of blood components remains a major challenge in transfusion medicine, particularly, platelet concentrates (PCs) due to the storage conditions that support bacterial proliferation. In this study, we develop a rapid, sensitive and specific real-time PCR protocol for bacterial screening of PCs. An internally controlled real-time PCR‑based method was optimized and validated with our proprietary 16S Universal PCR Master Mix (IBMP/Fiocruz), which targets a conserved region of the bacterial 16S rRNA gene. Nonspecific background DNA was completely eliminated by treating the PCR Master Mix with ethidium monoazide (EMA). A lower limit of detection was observed for 10 genome equivalents with an observed Ct value of 34±1.07 in calibration curve generated with 10-fold serial dilutions of E. coli DNA. The turnaround time for processing, including microbial DNA purification, was approximately 4 hours. The developed method showed a high sensitivity with no non-specific amplification and a lower time-to-detection than traditional microbiological methods, demonstrating it to be an efficient means of screening pre-transfusion PCs. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil. / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil. / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
| Subject | Bacterial Infections | pt_BR |
| Subject | Real-Time Polymerase Chain Reaction | pt_BR |
| Subject | Molecular Diagnostic Techniques | pt_BR |
| Subject | Blood Platelets | pt_BR |
| Subject in Spanish | Infecciones Bacterianas | pt_BR |
| Subject in Spanish | Reacción en Cadena en Tiempo Real de la Polimerasa | pt_BR |
| Subject in Spanish | Técnicas de Diagnóstico Molecular | pt_BR |
| Subject in Spanish | Plaquetas | pt_BR |
| Subject in French | Infections bactériennes | pt_BR |
| Subject in French | Réaction de polymérisation en chaine en temps réel | pt_BR |
| Subject in French | Techniques de diagnostic moléculaire | pt_BR |
| Subject in French | Plaquettes | pt_BR |
| DeCS | Infecções Bacterianas | pt_BR |
| DeCS | Reação em Cadeia da Polimerase em Tempo Real | pt_BR |
| DeCS | Técnicas de Diagnóstico Molecular | pt_BR |
| DeCS | Plaquetas | pt_BR |
