title: A rapid method for identifying Byssochlamys and Hamigera.
abstract: Heat-resistant fungi, genera Byssochlamys, Talaromyces, Neosartorya, and Hamigera, contribute significantly to the spoilage of heat-processed acidic foods, due to the formation of heat-resistant ascospores. Here, we first evaluated the differences in the beta-tubulin gene between Byssochlamys and Hamigera and developed specific primers to identify the Byssochlamys species fulva, nivea, and spectabilis, and Hamigera. Using primers designed for B. fulva and B. nivea (B1F/1R), specific PCR products were detected for B. fulva and B. nivea, as well as B. langunculariae and B. zollerniae, two closely related species. Similarly, the Pae4F/4R-1 and H2F/2R primers produced specific PCR products for B. spectabilis and Hamigera, respectively. Using these three primer sets, strains involved in acidic food spoilage and environmental contamination were not detected. The detection limits of all primer sets were 1 ng of DNA by PCR and 10 pg of DNA by nested PCR. Each PCR assay was specific, even if the sample was contaminated 1,000-fold by other fungal DNA. Thus, this method has proved to possess an extremely high degree of specificity.
authors.affiliation: Global R&D Safety Science, Kao Corporation, 2606 Akabane, Ichikai-machi, Haga-Gun, Tochigi 321-3497, Japan.
authors.forename: Motokazu
authors.initials: M
authors.lastname: Nakayama
authors.forename: Kouichi
authors.initials: K
authors.lastname: Hosoya
authors.forename: Tetsuhiro
authors.initials: T
authors.lastname: Matsuzawa
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Hiro
authors.forename: Ayumi
authors.initials: A
authors.lastname: Sako
authors.forename: Hajime
authors.initials: H
authors.lastname: Tokuda
authors.forename: Takashi
authors.initials: T
authors.lastname: Yaguchi
mesh_terms: D055323:Byssochlamys / Q000302:isolation & purification*; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis*; D032641:Eurotiales / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008969:Molecular Sequence Data; D055326:Neosartorya / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D013045:Species Specificity; D013172:Spores, Fungal; D032901:Talaromyces / Q000302:isolation & purification; D014404:Tubulin / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D014404:Tubulin