TY  - JOUR
PY  - 2019//
TI  - Rapid identification of Gloriosa superba and Colchicum autumnale by melting curve analysis: application to a suicide case involving massive ingestion of G. superba
JO  - International journal of legal medicine
A1  - Sakurada, Makoto
A1  - Yoshioka, Naoki
A1  - Kuse, Azumi
A1  - Nakagawa, Kanako
A1  - Morichika, Mai
A1  - Takahashi, Motonori
A1  - Kondo, Takeshi
A1  - Asano, Migiwa
A1  - Ueno, Yasuhiro
SP  - 1065
EP  - 1073
VL  - 133
IS  - 4
N2  - The plant species Gloriosa superba and Colchicum autumnale produce extremely poisonous colchicine as a major toxic metabolite. Almost all previous studies on colchicine poisoning have focused on drug analysis and clinical and pathological aspects. In this study, we developed a rapid, highly sensitive method to identify G. superba and C. autumnale. This method, which can distinguish between G. superba and C. autumnale using even minute amounts of plant material, is based on duplex real-time PCR in combination with melting curve analysis. To discriminate between the two genera of colchicine-containing plants, we designed new primer pairs targeting the region of the ycf15 gene, which is present in C. autumnale but not G. superba. By producing PCR amplicons with easily distinguishable melting temperatures, we were able to rapidly and accurately distinguish G. superba from C. autumnale. The new primer pairs generated no PCR amplicons from commercially available human DNA or various plant DNAs except for G. superba and C. autumnale. Sensitivity testing indicated that this assay can accurately detect less than 0.031 ng of DNA. Using our method in conjunction with colchicine drug analysis, we successfully identified G. superba in the stomach contents of a suicide victim who ingested massive quantities of a colchicine-containing plant. According to these results, duplex real-time PCR analysis is very appropriate for testing forensic samples, such as stomach contents harboring a variety of vegetables, and enables discrimination between G. superba and C. autumnale in forensic and emergency medical fields.<p />  <p>Language: en</p>
LA  - en
SN  - 0937-9827
UR  - http://dx.doi.org/10.1007/s00414-019-02060-x
ID  - ref1
ER  -