• Treffer 7 von 186
Zurück zur Trefferliste

A novel qPCR protocol to the specific detection and quantification of Hormoconis resinae in fuel systems

  • A novel qPCR protocol to the specific detection and quantification of Hormoconis resinae in fuel systems P.M. Martin-Sanchez1 , J. Toepel1, H.J. Kunte1, A.A. Gorbushina1, 2 1BAM Federal Institute for Materials Research and Testing, Department 4 Materials & Environment, Berlin, Germany 2 Free University of Berlin, Department of Earth Sciences & Biology, Chemistry & Pharmacy, Berlin, Germany Introduction. Microbial contamination of fuels is a phenomenon widely reported in the literature causing dramatic problems in fuel systems such as blockage of pipelines and filters, and corrosion of their metal components. A variety of microorganisms, fungi, yeasts and bacteria, can contaminate the distillate fuels. Between them, the filamentous fungus Hormoconis resinae, previously described as Cladosporium resinae and also known as "kerosene fungus" or "diesel bug", has traditionally been considered the main responsible of such troubles due to its great capacity to produce biomass andA novel qPCR protocol to the specific detection and quantification of Hormoconis resinae in fuel systems P.M. Martin-Sanchez1 , J. Toepel1, H.J. Kunte1, A.A. Gorbushina1, 2 1BAM Federal Institute for Materials Research and Testing, Department 4 Materials & Environment, Berlin, Germany 2 Free University of Berlin, Department of Earth Sciences & Biology, Chemistry & Pharmacy, Berlin, Germany Introduction. Microbial contamination of fuels is a phenomenon widely reported in the literature causing dramatic problems in fuel systems such as blockage of pipelines and filters, and corrosion of their metal components. A variety of microorganisms, fungi, yeasts and bacteria, can contaminate the distillate fuels. Between them, the filamentous fungus Hormoconis resinae, previously described as Cladosporium resinae and also known as "kerosene fungus" or "diesel bug", has traditionally been considered the main responsible of such troubles due to its great capacity to produce biomass and biofilms. Aim. The overarching goal of this study was to develop a novel real-time quantitative PCR (qPCR) method to detect and quantify the fungus H. resinae in samples from fuel systems. Materials and methods. Four specific primer sets targeting on two molecular markers, RNA polymerase II second largest subunit (RPB2) and internal transcribed spacers of rDNA (ITS), were designed using Primer Blast. The specificity of primer sets was evaluated by both conventional PCR and qPCR, analyzing the DNA extracts from H. resinae strains and other fungi and yeasts previously described in fuels as well as four taxonomically closest related species. The efficiency and sensitivity of these protocols were assessed by constructing the corresponding standard curves for each primer sets. Results. Two primer sets, Hr556F/Hr814R (RPB2) and Hr101F/Hr408R (ITS), were selected according to their remarkable specificity. Their standard curves showed a good correlation coefficient and efficiency. Additionally, the selected primer pairs showed a high sensitivity detecting a tiny amount of H. resinae DNA, 10 pg for RPB2 primers and 0.1 pg for ITS primers. Conclusions. The established qPCR protocol is a reliable and useful tool to the specific detection and quantification of H. resinae. Its subsequent implementation to analyze samples of fuels or biofilms covering fuel systems will allow the early detection of fungal outbreaks in order to apply the appropriated control procedures. Moreover, these analyses will determine the current incidence of this species in the cases of biological fuel contamination. Keywords. Hormoconis resinae, real-time qPCR, diesel bug, kerosene funguszeige mehrzeige weniger

Metadaten exportieren

Weitere Dienste

Teilen auf Twitter Suche bei Google Scholar
Metadaten
Autoren/innen:Pedro Maria Martin-Sanchez
Koautoren/innen:Anna Gorbushina, Jörg Toepel, Hans-Jörg Kunte
Dokumenttyp:Vortrag
Veröffentlichungsform:Präsentation
Sprache:Deutsch
Titel des übergeordneten Werkes (Deutsch):XVI International Biodeterioration and Biodegradation Symposium; International Biodeterioration & Biodegradation Society
Jahr der Erstveröffentlichung:2014
Organisationseinheit der BAM:4 Material und Umwelt
4 Material und Umwelt / 4.1 Biologische Materialschädigung und Referenzorganismen
Veranstaltung:XVI International Biodeterioration and Biodegradation Symposium; International Biodeterioration & Biodegradation Society
Veranstaltungsort:Lodz, Poland
Beginndatum der Veranstaltung:2014-09-03
Enddatum der Veranstaltung:2014-09-05
Verfügbarkeit des Dokuments:Weder Datei noch physisches Exemplar vorhanden ("No Access")
Datum der Freischaltung:20.02.2016
Referierte Publikation:Nein