A Case Study: What Doses of Amanita phalloides and Amatoxins Are Lethal to Humans?

Yılmaz, İsmail; Ermış, Fatih; Akata, İlgaz; Kaya, Ertuğrul

doi:10.1016/j.wem.2015.08.002

Cited by 37 publications

(19 citation statements)

References 25 publications

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“…Following the screening of the fusion plates, there were 14 positive cultures (optical density > 0. 7), of which 12 cultures exhibited substantial signal reduction (optical density decreased by 0.5 or greater) in the presence of 100 ng mL −1 α-AMA in cELISA ( Figure 2). Only two (9C12 and 9G3) of these grew stably, and were cloned multiple times until every well of the cell culture plate with cell growth elicited a positive indirect ELISA response to the coating antigen, a periodate-oxidized form of α-AMA conjugated to bovine serum albumin (PERI-AMA-BSA).…”

Section: Monoclonal Antibody Productionmentioning

confidence: 99%

“…The principle toxins responsible for the poisonings are the bicyclic octapeptides known as AMAs, most notably α-amanitin (α-AMA) and β-amanitin (β-AMA), and possibly γ-amanitin (γ-AMA) ( Figure 1). In mice, the LD 50 for α-AMA is 0.1 mg kg −1 [6] and, to humans, a dose of 0.3 mg kg −1 is severely toxic [7]. AMAs are potent inhibitors of RNA polymerase II, with bioactivity resistant to heat, cold, or acid inactivation.…”

Section: Introductionmentioning

confidence: 99%

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A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins

Bever

Hnasko

Cheng

et al. 2019

Toxins

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Amatoxins (AMAs) are lethal toxins found in a variety of mushroom species. Detection methods are needed to determine the occurrence of AMAs in mushroom species suspected in mushroom poisonings. In this manuscript, we report the generation of novel monoclonal antibodies (mAbs, AMA9G3 and AMA9C12) and the development of a competitive, enzyme-linked immunosorbent assay (cELISA) that is sensitive at 1 ng mL −1 and shows selectivity for α-amanitin (α-AMA) and γ-amanitin (γ-AMA), and less for β-amanitin (β-AMA). In order to decrease the overall time needed for analysis, the extraction procedure for mushrooms was also simplified. A rapid (1 min) extraction procedure of AMAs using solvents as simple as water alone was successfully demonstrated using Amanita mushrooms. Together, the extraction method and the mAb-based ELISA represent a simple and rapid method that readily detects AMAs extracted from mushroom samples. Key Contribution:Highly sensitive and selective monoclonal antibodies were developed to detect amatoxins. A rapid and simple extraction of amatoxins from mushroom specimens was demonstrated.

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Section: Monoclonal Antibody Productionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins

Bever

Hnasko

Cheng

et al. 2019

Toxins

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“…Amatoxins are highly resistant to degradation, and on the cellular level they inhibit transcription by binding to RNA polymerase II. As little as 0.1 mg/kg body weight of amatoxins may cause death [3,4], and this amount can be found in a single Amanita phalloides.…”

Section: Introductionmentioning

confidence: 99%

“…Toxins 2020, 12, x FOR PEER REVIEW 2 of 9 body weight of amatoxins may cause death [3,4], and this amount can be found in a single Amanita phalloides.…”

Section: Introductionmentioning

confidence: 99%

Rapid, Sensitive, and Accurate Point-of-Care Detection of Lethal Amatoxins in Urine

Bever

Swanson

Hamelin

et al. 2020

Toxins

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Globally, mushroom poisonings cause about 100 human deaths each year, with thousands of people requiring medical assistance. Dogs are also susceptible to mushroom poisonings and require medical assistance. Cyclopeptides, and more specifically amanitins (or amatoxins, here), are the mushroom poison that causes the majority of these deaths. Current methods (predominantly chromatographic, as well as antibody-based) of detecting amatoxins are time-consuming and require expensive equipment. In this work, we demonstrate the utility of the lateral flow immunoassay (LFIA) for the rapid detection of amatoxins in urine samples. The LFIA detects as little as 10 ng/mL of α-amanitin (α-AMA) or γ-AMA, and 100 ng/mL of β-AMA in urine matrices. To demonstrate application of this LFIA for urine analysis, this study examined fortified human urine samples and urine collected from exposed dogs. Urine is sampled directly without the need for any pretreatment, detection from urine is completed in 10 min, and the results are read by eye, without the need for specialized equipment. Analysis of both fortified human urine samples and urine samples collected from intoxicated dogs using the LFIA correlated well with liquid chromatography–mass spectrometry (LC-MS) methods.

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“…Fungi in the genus Amanita form ectomycorrhizal in association with plants, where they stimulate the growth and defense of the host organism [1,2]. However, Amanita mushrooms can also produce some of the most toxic compounds in the world [3][4][5]. Every year mushroom poisoning caused by mushroom con- sumption is responsible for fatal accidents, in particular in China [6].…”

Section: Introductionmentioning

confidence: 99%

Selection and characterization of DNA aptamer specially targeting α-amanitin in wild mushrooms

Han¹,

Xia²,

Jing³

et al. 2018

SDRP-JFST

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α-amanitin is a polypeptide isolated from the fruiting body of Amanita exitialis. It is the main toxin in wild mushrooms and toxic, often lethal, in animal and humans. In this study, the artificial nucleic acid aptamers targeting α-amanitin were screened by Systematic Evolution of Ligands by Exponential Enrichment (SELEX) in vitro, in order to develop an analytical tool for α-amanitin detection. The specificity of aptamer H06 with α-amanitin was confirmed using Enzyme-Linked OligoNucleotide Assay (ELONA) and Dot blot, and no non-specific was observed. Based on the ELONA platform, the minimum detectable concentration of aptamer H06 for α-amanitin was 8 ng/mL. The circular dichroism (CD) spectroscopy experiment indicated aptamer H06 forms a stem-loop and intramolecular G-quadruplex and it can stable exist in binding buffer and PBS buffer. Moreover , the affinity test showed a strong binding force between α-amanitin and the aptamer H06, with the dissociation constant (KD) of 37.5±5.135 nM. And the accurary of the ELONA assay based on aptame H06 was demonstrated in real mushroom samples. In summary, our data could demonstrate a possibility of the development of apta-based diagnostic platform and detection method for α-amanitin.

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A Case Study: What Doses of Amanita phalloides and Amatoxins Are Lethal to Humans?

Cited by 37 publications

References 25 publications

A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins

A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins

Rapid, Sensitive, and Accurate Point-of-Care Detection of Lethal Amatoxins in Urine

Selection and characterization of DNA aptamer specially targeting α-amanitin in wild mushrooms

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