Abstract
BackgroundA large proportion of neonates are treated for presumed bacterial sepsis with broad spectrum antibiotics even though their blood cultures subsequently show no growth. This study aimed to investigate PCR-based methods to identify pathogens not detected by conventional culture.MethodsWhole blood samples of 208 neonates with suspected early onset sepsis were tested using a panel of multiplexed bacterial PCRs targeting Streptococcus pneumoniae, Streptococcus agalactiae (GBS), Staphylococcus aureus, Streptococcus pyogenes (GAS), Enterobacteriaceae, Enterococcus faecalis, Enterococcus faecium, Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium, a 16S rRNA gene broad-range PCR and a multiplexed PCR for Candida spp.ResultsTwo-hundred and eight samples were processed. In five of those samples, organisms were detected by conventional culture; all of those were also identified by PCR. PCR detected bacteria in 91 (45%) of the 203 samples that did not show bacterial growth in culture. S. aureus, Enterobacteriaceae and S. pneumoniae were the most frequently detected pathogens. A higher bacterial load detected by PCR was correlated positively with the number of clinical signs at presentation.ConclusionReal-time PCR has the potential to be a valuable additional tool for the diagnosis of neonatal sepsis.
Highlights
Treatment for presumed sepsis is common in the neonatal period
Whole blood samples of 208 neonates with suspected early onset sepsis were tested using a panel of multiplexed bacterial PCRs targeting Streptococcus pneumoniae, Streptococcus agalactiae (GBS), Staphylococcus aureus, Streptococcus pyogenes (GAS), Enterobacteriaceae, Enterococcus faecalis, Enterococcus faecium, Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium, a 16S rRNA gene broadrange PCR and a multiplexed PCR for Candida spp
The first-choice antibiotic regimen for empirical treatment of suspected Early onset sepsis (EOS) recommended by the National Institute for Health a and Care Excellence is bencylpenicillin with gentamicin
Summary
Treatment for presumed sepsis is common in the neonatal period. Early onset sepsis (EOS) is defined as infection occurring within the first 48–72 hours of life, most often acquired shortly before or during delivery. Because a negative blood culture cannot exclude infection, antibiotic therapy is often continued beyond 48 hours despite negative blood cultures, especially, but not always, when there are clinical signs and laboratory markers consistent with infection [4]. This increases antibiotic usage, which in turn increases the risk of development of antibiotic resistant pathogens [5,6,7,8], as well as potential immune dysregulation in childhood, as a result of disruptions in nascent gut microbiome [9]. This study aimed to investigate PCR-based methods to identify pathogens not detected by conventional culture
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