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Real‐time PCR of the 16S‐rRNA gene in the diagnosis of neonatal bacteraemia
Author(s) -
Ohlin Andreas,
Bäckman Anders,
Björkqvist Maria,
Mölling Paula,
Jurstrand Margaretha,
Schollin Jens
Publication year - 2008
Publication title -
acta pædiatrica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 115
eISSN - 1651-2227
pISSN - 0803-5253
DOI - 10.1111/j.1651-2227.2008.00924.x
Subject(s) - medicine , blood culture , polymerase chain reaction , 16s ribosomal rna , sepsis , real time polymerase chain reaction , neonatal sepsis , coagulase , bacteremia , staphylococcus , cons , predictive value , microbiological culture , microbiology and biotechnology , staphylococcus aureus , positive predicative value , bacteria , gene , biology , antibiotics , genetics , computer science , programming language
Objective: To evaluate a real‐time PCR assay for the diagnosis of neonatal bacteraemia. Patients and methods: 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. Results: 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). Conclusion: 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.