Rapid identification of Gloriosa superba and Colchicum autumnale by melting curve analysis: application to a suicide case involving massive ingestion of G. superba
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 sensitiv...
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Published in | International journal of legal medicine Vol. 133; no. 4; pp. 1065 - 1073 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Berlin/Heidelberg
Springer Berlin Heidelberg
01.07.2019
Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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ISSN: | 0937-9827 1437-1596 |
DOI: | 10.1007/s00414-019-02060-x |