RT Journal Article
SR Electronic
T1 Rapid quantification of bacteria associated to ventilator associated pneumonia in endotracheal aspirate specimens
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP P2492
VO 44
IS Suppl 58
A1 Bruno François
A1 Sandrine Prudent
A1 Olivier Barraud
A1 Morgane Lacroix
A1 Marc Clavel
A1 Marie-Astrid Jestin
A1 Marc Rodrigue
A1 Marie-Cécile Ploy
A1 Alexandre Pachot
A1 Javier Yugueros-Marcos
A1 Virginie Moucadel
YR 2014
UL http://erj.ersjournals.com/content/44/Suppl_58/P2492.abstract
AB Ventilator-Associated Pneumonia (VAP) is one of the most serious infection in Intensive Care Unit. Today, its diagnosis relies mainly on conventional microbiological reports that are provided to the clinicians 24-72h after the sampling. Because an earlier diagnosis may reduce patient mortality and abusive use of antibiotics, we optimized a molecular biology-based method to quantify the DNA of bacteria frequently involved in VAP, directly from EndoTracheal Aspirates (ETA), in less than 3h30.First, a Sample Processing Control is spiked into each 400 μL specimen. Second, the sample is submitted to a liquefaction process and an enzymatic lysis, and DNA is extracted using the easyMAG® system. Then, qPCR are run and DNA is quantified using standard curves.Dynamic ranges for quantification were between ∼10^3 and ∼10^7 CFU/mL, depending on the bacterial target. The method was tested using ETA collected daily from mechanically ventilated patients (see patients 1 and 2, Figure 1) and we could follow the colonization with S. aureus over time. Indeed, the same trends were observed between the semi-quantitative culture and the ETA method.Further assessment is now needed to evaluate the medical validity of this method for the early diagnosis of VAP, on larger populations.