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Erschienen in: Forensic Toxicology 1/2021

04.11.2020 | Short Communication

Rapid identification of Colchicum autumnale based on loop-mediated isothermal amplification (LAMP) assay

verfasst von: Sae Misawa, Daigo Natsuhara, Yuka Kiba, Tadashi Yamamuro, Ryuichiro Suzuki, Takayuki Shibata, Masashi Kitamura

Erschienen in: Forensic Toxicology | Ausgabe 1/2021

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Abstract

Purpose

Colchicum autumnale (Colchicaceae) exhibits toxicity, and severe poisoning cases due to ingestion of this plant have been reported in Japan. Identifying the cause of poisoning is important for emergency medical treatment, and a rapid and simple detection technique is required for the identification of the cause of poisoning. In the present study, we developed a rapid and simple method for the detection of C. autumnale based on a loop-mediated isothermal amplification (LAMP) assay, which is 100- to 1000-fold higher than the conventional polymerase chain reaction method.

Methods

Specific LAMP primers for C. autumnale were designed based on the trnL-trnF intergenic spacer region. Using the LAMP primers, the LAMP assay was performed at an isothermal reaction temperature of 63 ℃.

Results

The LAMP reaction was shown to be specific and highly sensitive to C. autumnale, given that the assay can be used for 10 pg of purified DNA. Using a simple protocol for on-site detection, the entire procedure from pretreatment to evaluation required approximately 1 h. Moreover, the LAMP method using a microfluidic device detected multiple genes, including C. autumnale-specific DNA regions, at a time.

Conclusions

The currently proposed protocol exhibits good potential as a screening method for C. autumnale poisoning in emergency medical care.
Literatur
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Zurück zum Zitat Sakurada M, Yoshioka N, Kuse A, Nakagawa K, Morichika M, Takahashi M, Kondo T, Asano M, Ueno Y (2019) Rapid identification of Gloriosa superba and Colchicum autumnale by melting curve analysis: application to a suicide case involving massive ingestion of G. superba. Int J Legal Med 133:1065–1073. https://​doi.​org/​10.​1007/​s00414-019-02060-x ( open access article) CrossRefPubMed Sakurada M, Yoshioka N, Kuse A, Nakagawa K, Morichika M, Takahashi M, Kondo T, Asano M, Ueno Y (2019) Rapid identification of Gloriosa superba and Colchicum autumnale by melting curve analysis: application to a suicide case involving massive ingestion of G. superba. Int J Legal Med 133:1065–1073. https://​doi.​org/​10.​1007/​s00414-019-02060-x ( open access article) CrossRefPubMed
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Metadaten
Titel
Rapid identification of Colchicum autumnale based on loop-mediated isothermal amplification (LAMP) assay
verfasst von
Sae Misawa
Daigo Natsuhara
Yuka Kiba
Tadashi Yamamuro
Ryuichiro Suzuki
Takayuki Shibata
Masashi Kitamura
Publikationsdatum
04.11.2020
Verlag
Springer Singapore
Erschienen in
Forensic Toxicology / Ausgabe 1/2021
Print ISSN: 1860-8965
Elektronische ISSN: 1860-8973
DOI
https://doi.org/10.1007/s11419-020-00557-4

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