DNA testing of suspected cannabis samples with exceptional morphology using a simple detection kit
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CASE REPORT
DNA testing of suspected cannabis samples with exceptional morphology using a simple detection kit Tadashi Yamamuro1 · Yukie Iwai2 · Maho Kawamura2 · Akihiro Nakamoto2 · Hiroki Segawa1 · Kenji Kuwayama1 · Kenji Tsujikawa1 · Tatsuyuki Kanamori1 · Yuko T. Iwata1 Received: 24 June 2020 / Accepted: 15 September 2020 © Japanese Association of Forensic Toxicology 2020
Abstract Purpose Cannabis is the most abused drug in the world and molecular biology techniques for the analysis of cannabis have attracted attention in recent years. We report a case of three persons conspiring to possess cannabis-like leaf pieces for profit. We present the results of DNA testing by a simple kit on suspected cannabis samples with exceptionally shaped leaves. Methods DNA was obtained from the samples of cannabis leaves by a simple and rapid procedure using a cannabis DNA detection kit or commercially available purified plant DNA extraction kit. Undiluted or diluted DNA solution was used as templates for amplification of cannabis-specific sequences by triplex PCR. The presence or absence of PCR amplicon was visually observed by a DNA chromatography detection system to determine whether the sample was cannabis or not. Results When the DNA was extracted by the simple procedure and used without dilution, PCR failed to identify cannabis samples. However, when purified DNA was used, PCR amplification was successful in identifying all cannabis samples. Notably, in case of DNA extracted by the simple procedure, 10 to 50-fold dilution of the solution reduced PCR inhibition and PCR amplification was successful. Conclusions Although the quality of the DNA used as PCR template needs to be considered, simple DNA testing with a kit is very effective for identifying suspected cannabis samples with exceptional morphology. Keywords Cannabis sativa · Unusual form · DNA testing · DNA detection kit · PCR · DNA chromatography
Introduction Cannabis is the most abused drug in the world and is tightly regulated in many countries. The forensic identification of cannabis is generally performed by physical examination and chemical analysis [1, 2]. Physical examination is based on the morphological characteristics of the plant, Cannabis sativa. Macroscopically, the leaves have characteristic features such as being palmately composed of 3–11 leaflets with serrated margins [1, 3]. There are two known subspecies of cannabis, “indica” and “ruderalis”, which share similar characteristic leaf morphology as described above. * Tadashi Yamamuro [email protected] 1
National Research Institute of Police Science, 6‑3‑1, Kashiwanoha, Kashiwa, Chiba 277‑0882, Japan
Forensic Science Laboratory, Hiroshima Prefectural Police Headquarters, 2‑26‑3, Konan, Naka‑ku, Hiroshima, Hiroshima 730‑0825, Japan
2
The minor differences amongst the subspecies are as follows: “sativa” possess thin lanceolate leaflets, “indica” have thick oblanceolate leaflets, and “ruderalis” tend to have smaller leaves with fewer elliptic leaflets [3]. Microscopically, cannabis can
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