title: Evaluation of fungal aerosols using Temporal Temperature Gradient Electrophoresis (TTGE) and comparison with culture.
abstract: Information obtained from fungal air samples can assist in the assessment of health hazards and can be useful in proactive indoor air quality monitoring. The objective of the present study was to evaluate the PCR-TTGE technique for the analysis of fungal diversity in the air. Eleven air samples were collected in five different sites using the bioimpactor CIP 10-M (Arelco). After a 2 hours sampling period, the collection liquid was recovered for subsequent cultivation and PCR-TTGE. A set of three fungi-specific primers (Fungcont 1, Fungcont 2+GC and Fungcont 3) was designed for the partial amplification of the 18S rRNA gene. The amplification was obtained in a single reaction tube by a semi-nested PCR. For identification, the TTGE bands were extracted and sequenced. PCR-TTGE allowed the clear separation of amplicons corresponding to distinct fungal species (both Ascomycota and Basidiomycota) that may be encountered in air. The number of fungal taxa detected after culture was systematically higher than the number of taxa found using PCR-TTGE. However, few fungal species were detected by PCR-TTGE and not by cultivation, suggesting that the combination of these approaches may provide a better analysis of fungal diversity in air samples than either method alone.
authors.affiliation: INRA, AFSSA, ENVA, UPVM, UMR 956, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.
authors.forename: Adélaïde
authors.initials: A
authors.lastname: Nieguitsila
authors.forename: Manjula
authors.initials: M
authors.lastname: Deville
authors.forename: Taoufik
authors.initials: T
authors.lastname: Jamal
authors.forename: Lénaïg
authors.initials: L
authors.lastname: Halos
authors.forename: Madeleine
authors.initials: M
authors.lastname: Berthelemy
authors.forename: René
authors.initials: R
authors.lastname: Chermette
authors.forename: Sophie
authors.initials: S
authors.lastname: Latouche
authors.forename: Pascal
authors.initials: P
authors.lastname: Arné
authors.forename: Jacques
authors.initials: J
authors.lastname: Guillot
mesh_terms: D000336:Aerosols; D000391:Air Microbiology; D044822:Biodiversity; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005658:Fungi / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D006358:Hot Temperature; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D000336:Aerosols; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S