title: A novel qPCR protocol for the specific detection and quantification of the fuel-deteriorating fungus Hormoconis resinae.
abstract: A wide variety of fungi and bacteria are known to contaminate fuels and fuel systems. These microbial contaminants have been linked to fuel system fouling and corrosion. The fungus Hormoconis resinae, a common jet fuel contaminant, is used in this study as a model for developing innovative risk assessment methods. A novel qPCR protocol to detect and quantify H. resinae in, and together with, total fungal contamination of fuel systems is reported. Two primer sets, targeting the markers RPB2 and ITS, were selected for their remarkable specificity and sensitivity. These primers were successfully applied on fungal cultures and diesel samples demonstrating the validity and reliability of the established qPCR protocol. This novel tool allows clarification of the current role of H. resinae in fuel contamination cases, as well as providing a technique to detect fungal outbreaks in fuel systems. This tool can be expanded to other well-known fuel-deteriorating microorganisms.
authors.affiliation: a Department 4, Materials & Environment , Bundesanstalt für Materialforschung und -prüfung (BAM) , Berlin , Germany.
authors.forename: Pedro M
authors.initials: PM
authors.lastname: Martin-Sanchez
authors.affiliation: a Department 4, Materials & Environment , Bundesanstalt für Materialforschung und -prüfung (BAM) , Berlin , Germany.|b Department of Biology, Chemistry & Pharmacy and Department of Earth Sciences , Freie Universität Berlin , Berlin , Germany.
authors.forename: Anna A
authors.initials: AA
authors.lastname: Gorbushina
authors.affiliation: a Department 4, Materials & Environment , Bundesanstalt für Materialforschung und -prüfung (BAM) , Berlin , Germany.
authors.forename: Hans-Jörg
authors.initials: HJ
authors.lastname: Kunte
authors.affiliation: a Department 4, Materials & Environment , Bundesanstalt für Materialforschung und -prüfung (BAM) , Berlin , Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Toepel
mesh_terms: D001203:Ascomycota / Q000235:genetics / Q000302:isolation & purification*; D003343:Corrosion; D004271:DNA, Fungal / Q000235:genetics*; D016681:Genome, Fungal; D006838:Hydrocarbons / Q000032:analysis* / Q000592:standards; D007648:Kerosene / Q000382:microbiology*; D012319:RNA Polymerase II / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D018570:Risk Assessment
chemical_list: D004271:DNA, Fungal; D006838:Hydrocarbons; D007648:Kerosene; C029662:JP5 jet fuel; D012319:RNA Polymerase II
keywords: Real-time quantitative PCR; diesel biodeterioration; fouling; indicator; microbial contamination; microbiologically influenced corrosion