Development of a real-time PCR assay for the rapid detection of Acinetobacter baumannii from whole blood samples.
Acinetobacter baumannii is a multidrug-resistant pathogen associated with severe infections in hospitalized patients, including pneumonia, urinary and bloodstream infections. Rapid detection of A. baumannii infection is crucial for timely treatment of septicemic patients. The aim of the present study was to develop a specific marker for a quantitative polymerase chain reaction (PCR) assay for the detection of A. baumannii. The target gene chosen is the biofilm-associated protein (bap) gene, encoding a cell surface protein involved in biofilm formation. The assay is specific for A. baumannii, allowing its discrimination from different species of Acinetobacter and other clinically relevant bacterial pathogens. The assay is able to detect one genomic copy of A. baumannii, corresponding to 4 fg of purified DNA, and 20 colony-forming units/ml using DNA extracted from spiked whole blood samples.
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Eliana
E
De Gregorio
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Emanuela
E
Roscetto
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Vita Dora
VD
Iula
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Marianna
M
Martinucci
Department of Public Health, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Raffaele
R
Zarrilli
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Pier Paolo
PP
Di Nocera
Department of Molecular Medicine and Medical Biotechnologies, Federico II University, Via S. Pansini 5, 80131 Naples, Italy.
Maria Rosaria
MR
Catania
D000151:Acinetobacter Infections / Q000097:blood / Q000382:microbiology*; D040981:Acinetobacter baumannii / Q000235:genetics / Q000302:isolation & purification*; D001769:Blood / Q000382:microbiology*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
Bacterial bloodstream infection; Blood culture; Molecular diagnostics; Sepsis; TaqMan real-time PCR