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Development of a PCR-based tool for detecting immunologically relevant Alt a 1 and Alt a 1 homologue coding sequences
M F Gabriel1, I Postigo2, A Gutiérrez-Rodríguez2
1Department 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.
A new detection tool accurately identifies airborne Alt a 1 allergen exposure from various fungal species. This tool aids in assessing asthma risks and implementing public health measures.
Area of Science:
- Allergen research
- Molecular biology
- Public health
Background:
- Alt a 1 is a key allergen from Pleosporaceae fungi, linked to asthma.
- Accurate detection of airborne Alt a 1 is crucial for asthma management.
Purpose of the Study:
- Develop a highly specific detection tool for airborne Alt a 1.
- Identify fungal species producing airborne Alt a 1.
Main Methods:
- Designed primers (Alta1CF/Alta1CR) based on conserved Alt a 1 nucleotide regions.
- Utilized polymerase chain reaction (PCR) amplification and sequencing.
- Tested primers against cDNA from thirteen common fungal species.
Main Results:
- The Alta1CF/Alta1CR primer set detected Alt a 1 and Alt a 1-like genes in A. alternata, A. tenuissima, A. infectoria, U. botrytis, and S. botryosum.
- This system demonstrated high specificity and sensitivity, yielding a ~180 bp amplicon.
- Alternative primers (A21F/A21R) were specific only to A. alternata and A. tenuissima.
Conclusions:
- A rapid, sensitive, and specific PCR-based tool for Alt a 1 detection was developed.
- This tool can effectively assess Alt a 1 exposure levels.
- Findings support informed public health interventions for asthma prevention and control.
