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Background  Nosocomial candidaemia is associated with high mortality rates in critically ill paediatric patients; thus, the early detection and identification of the infectious agent is crucial for successful medical intervention. The PCR-based techniques have significantly increased the detection of  Candida  species in bloodstream infections. In this study, a multiplex nested PCR approach was developed for candidaemia detection in neonatal and paediatric intensive care patients.    Methods  DNA samples from the blood of 54 neonates and children hospitalised in intensive care units with suspected candidaemia were evaluated by multiplex nested PCR with specific primers designed to identify seven  Candida  species, and the results were compared with those obtained from blood cultures.    Results  The multiplex nested PCR had a detection limit of four  Candida  genomes/mL of blood for all  Candida  species. Blood cultures were positive in 14.8% of patients, whereas the multiplex nested PCR was positive in 24.0% of patients, including all culture-positive patients. The results obtained with the molecular technique were available within 24 hours, and the assay was able to identify  Candida  species with 100% of concordance with blood cultures. Additionally, the multiplex nested PCR detected dual candidaemia in three patients.    Conclusions  Our proposed PCR method may represent an effective tool for the detection and identification of  Candida  species in the context of candidaemia diagnosis in children, showing highly sensitive detection and the ability to identify the major species involved in this infection.

A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients