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Updated: Jan 17, 2026
Rapid High-throughput Species Identification of Botanical Material Using Direct Analysis in Real Time High Resolution Mass Spectrometry
Published on: October 2, 2016
Molecular identification of Amanita ibotengutake from patient vomit specimens in mushroom poisoning
Kosuke Ota1, Kengo Taira1, Ikuko Ito1
1Yamagata Prefectural Institute of Public Health, Yamagata, Japan.
Background:
Mushroom poisoning remains a global public health challenge, with morphological identification accuracy limited to 17%. This study aimed to demonstrate molecular identification from vomit specimens in a patient with mushroom poisoning.
Methods:
Following a mushroom poisoning incident in Yamagata Prefecture, Japan (September 2018), we collected four vomit specimens from a female patient. Deoxyribonucleic acid analysis targeting the internal transcribed spacer region using polymerase chain reaction and sequencing was performed on three of four specimens, while liquid chromatography-tandem mass spectrometry or toxin quantification was conducted on all four specimens. Species identification was conducted through sequence homology analysis.
Results:
Polymerase chain reaction amplification of the 185 base pair target sequence was successful from three vomit specimens collected 7 h post-ingestion, yielding identical sequences. Sequence analysis identified Amanita ibotengutake (100% homology), contradicting the initial morphological identification of Amanita pantherina. Liquid chromatography-tandem mass spectrometry detected a total of 0.053 mg ibotenic acid and 0.0043 mg muscimol in vomit specimens, consistent with the patient's neurological symptoms.
Discussions:
The discrepancy between morphological (Amanita pantherina) and molecular (Amanita ibotengutake) identification demonstrates limitations of conventional methods, particularly significant as these species were considered identical until 2002. Successful deoxyribonucleic acid amplification 7 h post-ingestion, despite technical challenges from gastric degradation, suggests clinical feasibility. This integrated molecular-toxicological approach strengthens scientific understanding of species-specific toxicities and contributes to improved epidemiological investigation of mushroom poisoning.
Conclusions:
We report the successful molecular identification from patient vomit specimens in a patient with mushroom poisoning. Although limited to a single case, this methodology may contribute to strengthening mushroom poisoning reports in the scientific literature to improve understanding of species-specific toxicities. Further validation in additional cases is needed for broader scientific applications.
Insights
Molecular identification of mushroom poisoning from vomit specimens successfully identified Amanita ibotengutake, correcting initial misidentification. This DNA-based method shows promise for accurate diagnosis and understanding species-specific toxicities in poisoning cases.
Area of Science:
- Forensic toxicology
- Molecular biology
- Mycology
Background:
- Mushroom poisoning is a global health issue with unreliable morphological identification (17% accuracy).
- Accurate species identification is crucial for understanding toxicity and patient management.
Purpose of the Study:
- To demonstrate the feasibility of molecular identification of mushroom poisoning from patient vomit specimens.
- To compare molecular identification with traditional morphological methods.
Main Methods:
- Collected four vomit specimens from a patient with suspected mushroom poisoning.
- Performed DNA analysis targeting the internal transcribed spacer region using PCR and sequencing.
- Utilized liquid chromatography-tandem mass spectrometry for toxin quantification.
Main Results:
- Identified *Amanita ibotengutake* with 100% sequence homology from three vomit specimens collected 7 hours post-ingestion.
- Detected ibotenic acid and muscimol in vomit specimens, consistent with neurological symptoms.
- Discrepancy noted between molecular identification (*A. ibotengutake*) and initial morphological identification (*Amanita pantherina*).
Conclusions:
- Molecular identification from vomit specimens is clinically feasible, even with gastric degradation.
- This integrated molecular-toxicological approach enhances understanding of mushroom poisoning and species-specific toxicities.
- Further validation in additional cases is required for broader application in scientific literature.
