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Real-time PCR of the 16S-rRNA gene in the diagnosis of neonatal bacteraemia
- Source :
- Acta Paediatrica. 97:1376-1380
- Publication Year :
- 2008
- Publisher :
- Wiley, 2008.
-
Abstract
- To evaluate a real-time PCR assay for the diagnosis of neonatal bacteraemia.Two hundred ninety-five plasma samples from 288 newborns with suspected neonatal sepsis were collected prospectively for the purpose of polymerase chain reaction (PCR)-based bacterial detection. A real-time PCR targeting the bacterial gene for 16S-rRNA gene combined with four specific probes designed to detect Gram-negative bacteria, Staphylococcus aureus and coagulase-negative staphylococci (CoNS) was developed. All samples positive in the universal PCR were further sequenced for bacterial identification.When applied to a material from 50 patients with positive blood culture and 245 patients with negative blood culture, the universal PCR showed a sensitivity of 42% (28-57), a specificity of 95% (92-97), a positive predictive value of 64% (45-80), and a negative predictive value of 89% (84-92) (95% confidence intervals in brackets).A new real-time PCR technique was for the first time applied to a well-defined prospectively and consecutively enrolled material of newborns with suspected sepsis, combining the benefits of real-time PCR with specific probes and sequencing. The method managed to detect bacteraemia with high specificity even though the sensitivity was low. Factors causing the low sensitivity are identified and further strategies to develop the method are described.
- Subjects :
- medicine.medical_specialty
Time Factors
Pcr assay
Bacteremia
Pilot Projects
Risk Assessment
Sensitivity and Specificity
law.invention
law
RNA, Ribosomal, 16S
Internal medicine
medicine
Humans
Gene
Polymerase chain reaction
Base Sequence
Plasma samples
Neonatal sepsis
Diagnostic Tests, Routine
Reverse Transcriptase Polymerase Chain Reaction
business.industry
Infant, Newborn
RNA Probes
General Medicine
16S ribosomal RNA
medicine.disease
Real-time polymerase chain reaction
Pediatrics, Perinatology and Child Health
Immunology
business
Subjects
Details
- ISSN :
- 16512227 and 08035253
- Volume :
- 97
- Database :
- OpenAIRE
- Journal :
- Acta Paediatrica
- Accession number :
- edsair.doi.dedup.....1a4e1131f1107885e5e5fd72634abb22