ArticleJournal of clinical microbiology2004
Two novel real-time reverse transcriptase PCR assays for rapid detection of bacterial contamination in platelet concentrates.
Article in Journal of clinical microbiology, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 54 citations in OpenAlex.
- An Internal Reference Control Duplex Real-Time Polymerase Chain Reaction Assay for Detecting Bacterial Contamination in Blood Products.PloS one · 2015Article
- Distribution of human-specific bacteroidales and fecal indicator bacteria in an urban watershed impacted by sewage pollution, determined using RNA- and DNA-based quantitative PCR assays.Applied and environmental microbiology · 2015Article
- Bacterial screening of platelet concentrates on day 2 and 3 with flow cytometry: the optimal sampling time point?Blood transfusion = Trasfusione del sangue · 2014Article
- Diagnostic methods for platelet bacteria screening: current status and developments.Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2014Review
- Emerging Pathogens - How Safe is Blood?Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2014Review
- Sensitive quantification of Clostridium difficile cells by reverse transcription-quantitative PCR targeting rRNA molecules.Applied and environmental microbiology · 2012Article
- BactQuant: an enhanced broad-coverage bacterial quantitative real-time PCR assay.BMC microbiology · 2012Article
- Implementation of Bacterial Detection Methods into Blood Donor Screening - Overview of Different Technologies.Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie · 2011Article
- Interactions between endocarditis-derived Streptococcus gallolyticus subsp. gallolyticus isolates and human endothelial cells.BMC microbiology · 2010Article
- Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice.Journal of clinical laboratory analysis · 2010Article
- Sensitive quantitative detection of commensal bacteria by rRNA-targeted reverse transcription-PCR.Applied and environmental microbiology · 2007Article
- Real-time quantitative broad-range PCR assay for detection of the 16S rRNA gene followed by sequencing for species identification.Journal of clinical microbiology · 2006Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The incidence of platelet bacterial contamination is approximately 1 per 2,000 units and has been acknowledged as the most frequent infectious risk from transfusion. In preliminary studies, the sterility of platelet concentrates (PCs) was tested with an automated bacterial blood culturing system and molecular genetic assays. Two real-time reverse transcriptase PCR (RT-PCR) assays performed in a LightCycler instrument were developed and compared regarding specificity and sensitivity by the use of different templates to detect the majority of the clinically important bacterial species in platelets. Primers and probes specific for the conserved regions of the eubacterial 23S rRNA gene or the groEL gene (encoding the 60-kDa heat shock protein Hsp60) were designed. During the development of the 23S rRNA RT-PCR, problems caused by the contamination of reagents with bacterial DNA were noted. Treatment with 8-methoxypsoralen and UV irradiation reduced the level of contaminating DNA. The sensitivity of the assays was greatly influenced by the enzyme system which was used. With rTth DNA polymerase in a one-enzyme system, we detected 500 CFU of Escherichia coli or Staphylococcus epidermidis/ml. With a two-enzyme system consisting of Moloney murine leukemia virus RT and Taq DNA polymerase, we detected 16 CFU/ml. With groEL mRNA as the target of RT-PCR under optimized conditions, we detected 125 CFU of E. coli/ml, and no problems with false-positive results caused by reagent contamination or a cross-reaction with human nucleic acids were found. Furthermore, the use of mRNA as an indicator of viability was demonstrated. Here we report the application of novel real-time RT-PCR assays for the detection of bacterial contamination of PCs that are appropriate for transfusion services.
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