Rapid Detection of Fungal Pathogens in Blood Cultures of Severely Burned Patients Using Panfungal PCR Assay

Background and Fungemia is a significant cause of morbidity and mortality among immunocompromised patients such as severe burns. Due to low sensitivity and long turnaround time of the traditional biphasic BHI blood culture to detect fungemia we aimed to detect fungal elements in blood culture of these patients suspected with invasive fungal disease (IFDs) using panfungal PCR assay. Four hundred blood cultures were obtained from 112 severely burned patients suspected with IFDs. DNA was extracted and a pair of fungal universal primers was used to amplify the 18SrRNA gene. The PCR product was sequenced and identified using the nucleotide Basic Local Alignment Search Tool (BLAST). 44(39.2%) Blood culture of 112 patients (mean age: 31.9±14.7 years) were positive. In PCR positive patients, mean burn size was 42.6±19 % of total body surface area and average hospital length of stay was 21.8 ±10.3 days. In three patients, same species of fungi were determined in both sequencing of PCR products and the classical procedures of blood culture. Randomly sequencing of 20 PCR products revealed Aspergillus fumigatus (n=11 55%), Candida tropicalis (n=5 25%), C. albicans (n=1 5%), C. parapsilosis (n=1 5%), Agaricomycotina (n=1 5%), and Penicillium (n=1 5%) as causative agents. Panfungal PCR assay on blood culture seems to be a promising method for rapid detection of fungi in blood culture of patients at risk for IFDs.