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Published on: June 25, 2020
Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Alternaria alternata
Xinyue Zhang1, Guojie Xu1, Huaqi Tang1
1Beijing University of Chinese Medicine, School of Chinese Pharmacy, No.6, Wangjing Central South Road, Chaoyang District, Beijing 100102, China.
Abstract:
Fungi of the Alternaria genus are associated with allergic diseases, with Alternaria alternata being one of the most prevalent species. A. alternata has been frequently reported as the etiologic agent of hypersensitivity pneumonitis, allergic rhinosinusitis, bronchial asthma, and other diseases. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay and a real-time PCR assay to detect low levels of A. alternata in herbal tea samples. The LAMP assay can detect as little as 3 pg/μL of A. alternata genomic DNA with high specificity. In addition, both the LAMP assay and the real-time PCR assay can be used for quantification of A. alternata. Although the newly developed LAMP assay is more rapid and specific in A. alternata identification, the real-time PCR assay is more precise in quantitation analysis.
Insights
This study presents new methods for detecting Alternaria alternata, a fungus linked to allergies. The developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays efficiently identify and quantify this common allergen in herbal teas.
Area of Science:
- Mycology
- Allergology
- Food Safety
Background:
- Fungi in the Alternaria genus are linked to various allergic diseases.
- Alternaria alternata is a prevalent species often implicated in hypersensitivity pneumonitis, allergic rhinosinusitis, and bronchial asthma.
- Accurate detection of A. alternata in food products like herbal teas is crucial for public health.
Purpose of the Study:
- To develop and validate sensitive molecular assays for detecting Alternaria alternata in herbal tea samples.
- To compare the performance of a novel loop-mediated isothermal amplification (LAMP) assay with a real-time PCR assay for A. alternata detection and quantification.
Main Methods:
- Development of a loop-mediated isothermal amplification (LAMP) assay.
- Development of a real-time PCR assay.
- Testing both assays for sensitivity, specificity, and quantification capabilities using varying concentrations of A. alternata genomic DNA.
Main Results:
- The LAMP assay demonstrated high specificity and could detect as little as 3 pg/μL of A. alternata genomic DNA.
- Both LAMP and real-time PCR assays were capable of quantifying A. alternata.
- The LAMP assay proved more rapid and specific for identification, while real-time PCR offered higher precision in quantification.
Conclusions:
- The developed LAMP assay provides a rapid and specific method for identifying Alternaria alternata in herbal teas.
- Real-time PCR offers more precise quantification of A. alternata.
- These assays enhance the ability to monitor and control A. alternata contamination in food products, contributing to allergy prevention.
