title: The Role of DNA Amplification and Cultural Growth in Complicated Acute Appendicitis.
abstract: Bacterial growth of peritoneal fluid specimens obtained during surgical procedures for acute appendicitis may be useful to optimize further antibiotic therapy in complicated cases. DNA amplification represents a fast technique to detect microbial sequences. We aimed to compare the potential of DNA amplificationversustraditional bacterial growth culture highlighting advantages and drawbacks in a surgical setting. Peritoneal fluid specimens were collected during surgery from 36 children who underwent appendectomy between May and December 2012. Real-time polymerase chain reaction (RT-PCR) and cultures were performed on each sample. RT-PCR showed an amplification of 16S in 18/36 samples,Escherichia coli(in 7 cases),Pseudomonas aeruginosa(3),Fusobacterium necrophorum(3),Adenovirus(2),E.coli(1),Klebsiella pneumoniae(1),Serratia marcescens/Enterobacter cloacae(1). Bacterial growth was instead observed only in four patients (3E.coliand 1P.aeruginosaandBacteroides ovatus). Preoperative C-reactive protein and inflammation degree, the most reliable indicators of bacterial translocation, were elevated as expected. DNA amplification was a quick and useful method to detect pathogens and it was even more valuable in detecting aggressive pathogens such as anaerobes, difficult to preserve in biological cultures; its drawbacks were the lack of biological growths and of antibiograms. In our pilot study RT-PCR and cultures did not influence the way patients were treated.
authors: Department of Pediatric Surgery, University of Florence and Children's University Hospital A. Meyer.
authors: Francesca
authors: F
authors: Tocchioni
authors: Department of Pediatric Surgery, University of Florence and Children's University Hospital A. Meyer.
authors: Chiara
authors: C
authors: Tani
authors: Department of Microbiology, Careggi Hospital , Florence.
authors: Laura
authors: L
authors: Bartolini
authors: Department of Clinical Immunology, University of Florence and Children's University Hospital A. Meyer , Italy.
authors: Maria
authors: M
authors: Moriondo
authors: Department of Clinical Immunology, University of Florence and Children's University Hospital A. Meyer , Italy.
authors: Francesco
authors: F
authors: Nieddu
authors: Department of Microbiology, Careggi Hospital , Florence.
authors: Patrizia
authors: P
authors: Pecile
authors: Department of Clinical Immunology, University of Florence and Children's University Hospital A. Meyer , Italy.
authors: Chiara
authors: C
authors: Azzari
authors: Department of Pediatric Surgery, University of Florence and Children's University Hospital A. Meyer.
authors: Antonio
authors: A
authors: Messineo
authors: Department of Pediatric Surgery, University of Florence and Children's University Hospital A. Meyer.
authors: Marco
authors: M
authors: Ghionzoli
keywords: Antibiotics therapy; Real-time-PCR; complicated appendicitis; microbiological culture; pathogens