title: Multiplex PCR allows rapid and accurate diagnosis of bloodstream infections in newborns and children with suspected sepsis.
abstract: Sepsis is a major health problem in newborns and children. Early detection of pathogens allows initiation of appropriate antimicrobial therapy that strongly correlates with positive outcomes. Multiplex PCR has the potential to rapidly identify bloodstream infections, compensating for the loss of blood culture sensitivity. In an Italian pediatric hospital, multiplex PCR (the LightCycler SeptiFast test) was compared to routine blood culture with 1,673 samples obtained from 803 children with suspected sepsis; clinical and laboratory information was used to determine the patient infection status. Excluding results attributable to contaminants, SeptiFast showed a sensitivity of 85.0% (95% confidence interval [CI] = 78.7 to 89.7%) and a specificity of 93.5% (95% CI = 92.1 to 94.7%) compared to blood culture. The rate of positive results was significantly higher with SeptiFast (14.6%) than blood culture (10.3%) (P < 0.0001), and the overall positivity rate was 16.1% when the results of both tests were combined. Staphylococcus aureus (11.6%), coagulase-negative staphylococci (CoNS) (29.6%), Pseudomonas aeruginosa (16.5%), and Klebsiella spp. (10.1%) were the most frequently detected. SeptiFast identified 97 additional isolates that blood culture failed to detect (24.7% P. aeruginosa, 23.7% CoNS, 14.4% Klebsiella spp., 14.4% Candida spp.). Among specimens taken from patients receiving antibiotic therapy, we also observed a significantly higher rate of positivity of SeptiFast than blood culture (14.1% versus 6.5%, respectively; P < 0.0001). On the contrary, contaminants were significantly more frequent among blood cultures than SeptiFast (n = 97 [5.8%] versus n = 26 [1.6%]), respectively; P < 0.0001). SeptiFast served as a highly valuable adjunct to conventional blood culture in children, adding diagnostic value and shortening the time to result (TTR) to 6 h.
authors.affiliation: Unit of Microbiology, Children's Hospital and Research Institute Bambino Gesù, Rome, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Lucignano
authors.forename: Stefania
authors.initials: S
authors.lastname: Ranno
authors.forename: Oliver
authors.initials: O
authors.lastname: Liesenfeld
authors.forename: Beatrice
authors.initials: B
authors.lastname: Pizzorno
authors.forename: Lorenza
authors.initials: L
authors.lastname: Putignani
authors.forename: Paola
authors.initials: P
authors.lastname: Bernaschi
authors.forename: Donato
authors.initials: D
authors.lastname: Menichella
mesh_terms: D000293:Adolescent; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D017848:Blood-Borne Pathogens / Q000302:isolation & purification*; D002175:Candida / Q000235:genetics / Q000302:isolation & purification; D002648:Child; D002675:Child, Preschool; D006761:Hospitals; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007558:Italy; D008828:Microbiological Techniques / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000175:diagnosis* / Q000209:etiology*