A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins
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Abstract:
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.Keywords:
Amanita
Mushroom poisoning
Mushroom poisoning
Edible mushroom
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Abstract Three human and three murine monoclonal antibodies were tested for their reactivity to tetanus toxin and toxoid and used to establish an enzyme immunoassay specific for tetanus toxin. The dissociation constants of the monoclonal antibodies were between 3.91 × 10 −9 and 8.48 × 10 −12 . Two human monoclonal antibodies recognized conformation determinants on the toxin, whereas the others reacted to the heavy chain. Only a combination of antibodies of the two species allowed the development of an enzyme immunoassay for the detection of tetanus toxin with a lower detection limit of 1.2 μg/l.
Toxoid
Clostridium tetani
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Mushroom poisoning
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The problem of mushroom intoxication has been brought up more often in both scientific journals and media, particularly in autumn – the period when mushroom hunting is most intense. Mushroom intoxication is mainly perceived as being associated with acute hepatic insufficiency. However, apart from strong hepatotoxic effects, some mushroom species are characterized by effects that lead to renal damage, sometimes irreversible in character. This article relates to the toxic mushroom species considered to be nephrotoxic and discusses different mechanisms and symptoms of kidney injury as well as methods of treatment
Mushroom poisoning
Nephrotoxicity
Edible mushroom
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Amanita
Mushroom poisoning
Agaricales
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To describe and analyze the clinical characteristics and outcome of amatoxin poisoning cases.We performed a retrospective cohort study of amatoxin poisoning cases from Ramathibodi Poison Center Toxic Exposure Surveillance System, from May 2013 to August 2015.There were 30 consultations with a total of 55 poisoning cases. Most cases were male and from the north-east region. Hepatitis, acute kidney injury, jaundice, and coagulopathy accounted for 74%, 46.3%, 44.7%, and 52.8% of the cases, respectively. Almost all of the patients were admitted to the hospital, and the median duration of hospital stay was found to be 4 days. Mortality rate was found to be 27.3%. Most patients (73%) received the treatment including multiple-dose activated charcoal (67.5%), intravenous N-acetylcysteine (87.5%), and benzylpenicillin (45%). In 60% of the cases, the treatment was initiated within 24 h after eating mushrooms. Exchange transfusion and liver transplantation were performed in one severe case. However, this patient died eventually. Because intravenous silybinin is not available in Thailand during the study period, 8 patients received oral silymarin instead. All 8 patients had hepatitis and were treated with high dosage of oral silymarin (5 patients with 4.48 g/day, 2 patients with 1.68 g/day, and 1 patient with 1.4 g/day) for a couple of days. One of these patients died as she received treatment very late; she was treated with silymarin at 1.68 g/day dosage. Thus, the fatality in oral silymarin treatment group was 12.5%. We performed the analysis between the dead and survival groups. We found that in hepatitis, initial and maximum serum aspartate transaminase, initial and maximum serum alanine transaminase, and acute kidney injury were significantly different between the two groups.Amanita mushroom poisoning caused high fatalities. Serum transaminase and creatinine were the factors associated with death. Treatment with oral high dose silymarin should be investigated further as one of the principal therapies in amatoxin poisoning.
Amanita
Mushroom poisoning
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Wild mushroom poisoning from the genus Amanita is a medical emergency, with Amanita phalloides being the most common offender. Patients may complain of nausea, vomiting, diarrhea and/or abdominal pain. If not aggressively treated, fulminant hepatic failure may develop within several days of ingestion. In this case report, a patient poisoned with Amanita bisporigera is described, along with the typical clinical presentation, patient outcomes, and treatment options for dealing with an Amanita mushroom poisoning.
Mushroom poisoning
Amanita
Amanita phalloides
Fulminant
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Amanita
Mushroom poisoning
Amanita phalloides
Liver disease
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Amanita
Mushroom poisoning
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The lack of experience and fundamental knowledge about mycology by some mushroomers is one of the leading causes of increasing occurrence of fatal mushroom poisonings. Mushroom intoxications are caused not only by poisonous mushrooms (true primary intoxications), but under certain conditions also by edible mushrooms (secondary intoxications, false intoxications, pseudo-intoxications). Apart from fresh mushrooms intoxications may result also from preserved mushrooms (sterilized in pickles, soured, dried, used for preparation of mushroom extracts, powders, etc.), which are used as garnish. (Tab. 1, Ref. 44.)
Mushroom poisoning
Edible mushroom
Agaricales
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