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Development of a Broad-Range 23S rDNA Real-Time PCR Assay for the Detection and Quantification of Pathogenic Bacteria in Human Whole Blood and Plasma Specimens
Joint Authors
Gaibani, Paolo
Mariconti, Mara
Bua, Gloria
Bonora, Sonia
Sassera, Davide
Sambri, Vittorio
Mulatto, Patrizia
Novati, Stefano
Bandi, Claudio
Landini, Maria Paola
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-03-11
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
Molecular methods are important tools in the diagnosis of bloodstream bacterial infections, in particular in patients treated with antimicrobial therapy, due to their quick turn-around time.
Here we describe a new broad-range real-time PCR targeting the 23S rDNA gene and capable to detect as low as 10 plasmid copies per reaction of targeted bacterial 23S rDNA gene.
Two commercially available DNA extraction kits were evaluated to assess their efficiency for the extraction of plasma and whole blood samples spiked with different amount of either Staphylococcus aureus or Escherichia coli, in order to find the optimal extraction method to be used.
Manual QIAmp extraction method with enzyme pre-treatment resulted the most sensitive for detection of bacterial load.
Sensitivity of this novel assay ranged between 10 and 103 CFU per PCR reaction for E.
coli and S.
aureus in human whole blood samples depending on the extraction methods used.
Analysis of plasma samples showed a 10- to 100-fold reduction of bacterial 23S rDNA in comparison to the corresponding whole blood specimens, thus indicating that whole blood is the preferential sample type to be used in this real-time PCR protocol.
Our results thus show that the 23S rDNA gene represents an optimal target for bacteria quantification in human whole blood.
American Psychological Association (APA)
Gaibani, Paolo& Mariconti, Mara& Bua, Gloria& Bonora, Sonia& Sassera, Davide& Landini, Maria Paola…[et al.]. 2013. Development of a Broad-Range 23S rDNA Real-Time PCR Assay for the Detection and Quantification of Pathogenic Bacteria in Human Whole Blood and Plasma Specimens. BioMed Research International،Vol. 2013, no. 2013, pp.1-8.
https://search.emarefa.net/detail/BIM-1030259
Modern Language Association (MLA)
Gaibani, Paolo…[et al.]. Development of a Broad-Range 23S rDNA Real-Time PCR Assay for the Detection and Quantification of Pathogenic Bacteria in Human Whole Blood and Plasma Specimens. BioMed Research International No. 2013 (2013), pp.1-8.
https://search.emarefa.net/detail/BIM-1030259
American Medical Association (AMA)
Gaibani, Paolo& Mariconti, Mara& Bua, Gloria& Bonora, Sonia& Sassera, Davide& Landini, Maria Paola…[et al.]. Development of a Broad-Range 23S rDNA Real-Time PCR Assay for the Detection and Quantification of Pathogenic Bacteria in Human Whole Blood and Plasma Specimens. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-8.
https://search.emarefa.net/detail/BIM-1030259
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1030259