title: Development of a PCR-based tool for detecting immunologically relevant Alt a 1 and Alt a 1 homologue coding sequences.
abstract: Alt a 1 has been recognized as the most important allergen produced by the Pleosporaceae family and is a risk factor for asthma development and/or exacerbation. The aim of this study was to develop a detection tool that is highly specific for species that produced airborne Alt a 1. Based on the highly conserved internal nucleotide region of the several Alt a 1 sequences that are available in GenBank, a pair of primers (Alta1CF/Alta1CR) was designed. A set of primers used by other authors for the production of recombinant Alt a 1 (A21F/A21R) was also tested. The molecular analyses were based on the polymerase chain reaction (PCR) amplification and sequencing of the cDNA that was obtained from thirteen common fungal species. The PCR system that utilized Alta1CF/Alta1CR was highly specific, sensitive, and was able to detect an amplicon of approximately 180 bp from Alt a 1 and Alt a 1-like encoding genes from A. alternata, A. tenuissima, A. infectoria, U. botrytis, and S. botryosum. In contrast, the A21F/A21R primers were specific for the very close taxonomically related species A. alternata and A. tenuissima. Thus, this rapid, sensitive, and specific detection tool can be used to assess Alt a 1 exposure levels and to inform the implementation of the appropriate public health measures.
authors: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and Laboratory of Parasitology and Allergy, Lascaray Research Centre, University of the Basque Country, Vitoria, Spain Department of Chemistry and CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal.
authors: M F
authors: MF
authors: Gabriel
authors: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and Laboratory of Parasitology and Allergy, Lascaray Research Centre, University of the Basque Country, Vitoria, Spain.
authors: I
authors: I
authors: Postigo
authors: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and Laboratory of Parasitology and Allergy, Lascaray Research Centre, University of the Basque Country, Vitoria, Spain.
authors: A
authors: A
authors: Gutiérrez-Rodríguez
authors: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and Laboratory of Parasitology and Allergy, Lascaray Research Centre, University of the Basque Country, Vitoria, Spain.
authors: E
authors: E
authors: Suñén
authors: Department of Chemistry and CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal.
authors: C T
authors: CT
authors: Tomaz
authors: Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy and Laboratory of Parasitology and Allergy, Lascaray Research Centre, University of the Basque Country, Vitoria, Spain jorge.martinez@ehu.es.
authors: J
authors: J
authors: Martínez
mesh_terms: D000946:Antigens, Fungal / Q000235:genetics*; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D004784:Environmental Monitoring; D005658:Fungi / Q000145:classification / Q000235:genetics*; D016533:Mycological Typing Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D000946:Antigens, Fungal; D004271:DNA, Fungal
keywords: Alt a 1; Alt a 1 homologues; Alternaria alternata; allergen monitoring; amplification