The role of DNA amplification and cultural growth in complicated acute appendicitis

• Main Authors: Francesca Tocchioni, Chiara Tani, Laura Bartolini, Maria Moriondo, Francesco Nieddu, Patrizia Pecile, Chiara Azzari, Antonio Messineo, Marco Ghionzoli
• Format: Article
• Language: English
• Published: MDPI AG 2016-09-01
• Series: Pediatric Reports
• Subjects: Antibiotics therapy; complicated appendicitis; microbiological culture; pathogens; Real-time-PCR.
• Online Access: http://www.pagepress.org/journals/index.php/pr/article/view/6487
• ISSN: 2036-749X; 2036-7503

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 amplification versus traditional 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, <em>Escherichia coli</em> (in 7 cases), <em>Pseudomonas aeruginosa</em> (3), <em>Fusobacterium necrophorum</em> (3), <em>Adenovirus</em> (2), <em>E.coli</em> (1), <em>Klebsiella pneumoniae</em> (1), <em>Serratia marcescens/Enterobacter cloacae</em> (1). Bacterial growth was instead observed only in four patients (3 <em>E.coli</em> and 1 <em>P.aeruginosa</em> and <em>Bacteroides ovatus</em>). 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.