A radioimmunoassay for amanitin

Faulstich, Heinz; Trischmann, Heinrich; Zobeley, Suse

doi:10.1016/0014-5793(75)81116-1

Cited by 43 publications

(12 citation statements)

References 5 publications

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“…For mushroom analysis, instrumental methods (e.g., liquid chromatographymass spectrometry (LC-MS)) are highly sensitive and selective, but require extensive sample pre-treatment and expensive equipment [23][24][25][26]. Immunoassays (e.g., enzyme-linked immunosorbent assays, ELISAs) are sensitive and selective, but still require specialized reagents and equipment, and take a few hours to perform [27][28][29][30][31][32]. However, these same immunoreagents used in an ELISA can be transferable to a lateral flow immunoassay (LFIA) format, which often significantly reduces the assay time and the need for specialized equipment.…”

Section: Plos Onementioning

confidence: 99%

Lateral flow immunoassay (LFIA) for the detection of lethal amatoxins from mushrooms

et al. 2020

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The mushroom poison that causes the most deaths is the class of toxins known as amatoxins. Current methods to sensitively and selectively detect these toxins are limited by the need for expensive equipment, or they lack accuracy due to cross-reactivity with other chemicals found in mushrooms. In this work, we report the development of a competition-based lateral flow immunoassay (LFIA) for the rapid, portable, selective, and sensitive detection of amatoxins. Our assay clearly indicates the presence of 10 ng/mL of α-AMA or γ-AMA and the method including extraction and detection can be completed in approximately 10 minutes. The test can be easily read by eye and has a presumed shelf-life of at least 1 year. From testing 110 wild mushrooms, the LFIA identified 6 out of 6 species that were known to contain amatoxins. Other poisonous mushrooms known not to contain amatoxins tested negative by LFIA. This LFIA can be used to quickly identify amatoxin-containing mushrooms.

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Section: Plos Onementioning

confidence: 99%

Lateral flow immunoassay (LFIA) for the detection of lethal amatoxins from mushrooms

et al. 2020

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“…Current methods for quantifying amatoxins include high performance liquid chromatography (HPLC) with detection by either ultraviolet (UV) or mass spectrometry (MS) [ 3 , 6 ], thin-layer chromatography (TLC) [ 7 ], capillary zone electrophoresis [ 8 ], or immunoassay [ 9 , 10 , 11 ]. The most sensitive HPLC method with a UV detector provides a detection limit of 10 µg/L of extraction buffer (corresponding to 0.5 ng/g mushroom tissue) [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

A New Conjugation Method Used for the Development of an Immunoassay for the Detection of Amanitin, a Deadly Mushroom Toxin

Bever

Barnych

Hnasko

et al. 2018

Toxins

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One of the deadliest mushrooms is the death cap mushroom, Amanita phalloides. The most toxic constituent is α-amanitin, a bicyclic octapeptide, which damages the liver and kidneys. To develop a new tool for detecting this toxin, polyclonal antibodies were generated and characterized. Both α- and β-amanitin were coupled to carrier proteins through four different linking chemistries, one of which has never before been described. These conjugates were evaluated for their effectiveness in generating antibodies specific for the free toxin, as well as their utility in formatting heterogeneous assays with high sensitivity. Ultimately, these efforts yielded a newly described conjugation procedure utilizing periodate oxidation followed by reductive amination that successfully resulted in generating sensitive immunoassays (limit of detection (LOD), ~1.0 µg/L). The assays were characterized for their selectivity and were found to equally detect α-, β-, and γ-amanitin, and not cross-react with other toxins tested. Toxin detection in mushrooms was possible using a simple sample preparation method. This enzyme-linked immunosorbent assay (ELISA) is a simple and fast test, and readily detects amatoxins extracted from A. phalloides.

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“…Although several methods have been described to determine amatoxins and/or phallotoxins in biological fluids, including radioimmunoassay (RIA) [ 11 , 12 , 13 ], enzyme-linked immunosorbent assay (ELISA) [ 14 , 15 ], capillary zone electrophoresis (CZE) [ 16 , 17 ], and liquid chromatography (LC) combined with ultraviolet (UV) [ 7 , 18 , 19 ] or electrochemical detection (ECD) [ 20 , 21 , 22 ], various problems, such as false positives, unstable reproducibility, poor confirmatory ability and laborious procedures, have limited their application in practice [ 19 , 23 ]. Methods combining LC with mass spectrometry (MS), such as triple quadrupole tandem MS (MS/MS) [ 24 , 25 , 26 , 27 , 28 ], time-of-flight (TOF) MS [ 29 ] and matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS) [ 30 ], have been reported for the detection of amatoxins and phallotoxins.…”

Section: Introductionmentioning

confidence: 99%

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

Zhang

Zhao

et al. 2016

Toxins

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Amatoxins and phallotoxins are toxic cyclopeptides found in the genus Amanita and are among the predominant causes of fatal food poisoning in China. In the treatment of Amanita mushroom poisoning, an early and definite diagnosis is necessary for a successful outcome, which has prompted the development of protocols for the fast and confirmatory determination of amatoxins and phallotoxins in human biological fluids. For this purpose, a simple, rapid and sensitive multiresidue UPLC-MS/MS method for the simultaneous determination of α-amanitin, β-amanitin, γ-amanitin, phalloidin (PHD) and phallacidin (PCD) in human plasma, serum and urine was developed and validated. The diluted plasma, serum and urine samples were directly purified with a novel PRiME technique on a 96-well μElution plate platform, which allowed high-throughput sample processing and low reagent consumption. After purification, a UPLC-MS/MS analysis was performed using positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. This method fulfilled the requirements of a validation test, with good results for the limit of detection (LOD), lower limit of quantification (LLOQ), accuracy, intra- and inter-assay precision, recovery and matrix effects. All of the analytes were confirmed and quantified in authentic plasma, serum and urine samples obtained from cases of poisoning using this method. Using the PRiME μElution technique for quantification reduces labor and time costs and represents a suitable method for routine toxicological and clinical emergency analysis.

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A radioimmunoassay for amanitin

Cited by 43 publications

References 5 publications

Lateral flow immunoassay (LFIA) for the detection of lethal amatoxins from mushrooms

Lateral flow immunoassay (LFIA) for the detection of lethal amatoxins from mushrooms

A New Conjugation Method Used for the Development of an Immunoassay for the Detection of Amanitin, a Deadly Mushroom Toxin

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

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