Universal identification of lethal amanitas by using Hyperbranched rolling circle amplification based on α-amanitin gene sequences
| Autor: | Tao Luo, Ping Zhang, Fengxia Fan, Zhengmi He, Zuohong Chen |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Amanita
Amanitins Loop-mediated isothermal amplification Mushroom Poisoning Polymerase Chain Reaction Sensitivity and Specificity 01 natural sciences Lepiota Analytical Chemistry law.invention 0404 agricultural biotechnology Limit of Detection law parasitic diseases Gene Alpha-Amanitin Amanitin biology 010401 analytical chemistry 04 agricultural and veterinary sciences General Medicine biology.organism_classification 040401 food science Molecular biology 0104 chemical sciences Rolling circle replication Recombinant DNA Galerina Agaricales Nucleic Acid Amplification Techniques Food Science |
| Zdroj: | Food Chemistry. 298:125031 |
| ISSN: | 0308-8146 |
| Popis: | Hyperbranched rolling circle amplification (HRCA) with a padlock probe (PLP) targeting the α-amanitin (α-AMA) gene, as a screening tool for the universal identification of lethal amanitas, was established in this study. With the isothermal HRCA assay, all of the lethal Amanita species tested from Phalloideae (10) were positive, while the non-Phalloideae Amanita species (15) and three amanitin-containing Lepiota and Galerina species were negative. Furthermore, the PLP based on α-AMA sequences from lethal Amanita species was effective for Amanita α-AMA, but not Amanita β-AMA or non-Amanita α-AMA. HRCA sensitivity was 100-fold higher than conventional PCR with a detection limit of 100 copies (recombinant plasmid containing α-AMA), and 0.2% lethal amanitas could be detected in dry mushroom blends. The HRCA method presented provided a rapid, specific, sensitive and low-cost identification tool for lethal amanitas. |
| Databáze: | OpenAIRE |
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