Development of a similarity discrimination method for cannabis samples using Inter-Simple Sequence Repeat-PCR.

IF 2.1 4区医学 Q2 TOXICOLOGY

Forensic Toxicology Pub Date : 2026-06-16 DOI:10.1007/s11419-026-00775-2

Tadashi Yamamuro, Yuki Okada, Hiroki Segawa, Kenji Kuwayama, Kenji Tsujikawa, Tatsuyuki Kanamori, Yuko T Iwata

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引用次数: 0

Abstract

Purpose: In forensic cannabis investigations, it is important not only to prove that seized materials are cannabis or not but also to clarify the relationship between cannabis samples. This study aimed to establish a method for evaluating the similarity of the genetic background of cannabis samples using Inter-Simple Sequence Repeat (ISSR)-PCR analysis, a variant of DNA testing.

Methods: Forty-eight cannabis samples from 24 strains were collected. ISSR-PCR was performed using 10 ISSR primers, and the resulting band patterns were used for grouping. After analyzing the characteristics of each primer using a binary classification method, two numerical indicators, the similarity score and total likelihood ratio, were calculated to comprehensively evaluate the similarity between samples and assess their discrimination performance.

Results: Cannabis samples were grouped into band patterns using ISSR-PCR, but no single primer classified all strains. The "similarity score" (0-10), based on primers showing the same band pattern across 10 ISSR-PCRs, averaged 1.69 between different strains, 4.34 between individuals of the identical strain, and 9.48 for the identical individual. The mean "total likelihood ratio" calculated from the positive and negative likelihood ratios for binary classification using 10 primers was 1.28 × 10⁰, 1.11 × 10⁵, and 9.79 × 10⁹, respectively. Each indicator demonstrated high discrimination performance in evaluating genetic similarity.

Conclusions: This study established numerical criteria for estimating the genetic similarity between unknown cannabis samples based on ISSR-PCR results using 10 primers.

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利用简单序列重复pcr建立大麻样品的相似性鉴别方法。

目的：在大麻法医调查中，重要的是不仅要证明缉获的材料是否是大麻，而且要澄清大麻样品之间的关系。本研究旨在利用DNA检测的一种变体——ISSR -PCR分析，建立一种评估大麻样本遗传背景相似性的方法。方法：采集24个大麻品系48份大麻样品。用10个ISSR引物进行ISSR- pcr，并将所得带型进行分组。采用二值分类法分析每个引物的特征后，计算相似度得分和总似然比两个数值指标，综合评价样本之间的相似度，评价其鉴别性能。结果：用ISSR-PCR方法将大麻样品分组成带型，但没有单一引物对所有菌株进行分类。在10条issr - pcr序列中，以具有相同条带模式的引物为基础的“相似性得分”（0-10），不同菌株之间平均为1.69分，同一菌株个体之间平均为4.34分，同一个体平均为9.48分。使用10个引物进行二元分类的正似然比和负似然比计算得出的平均“总似然比”分别为1.28 × 10⁰、1.11 × 10 5和9.79 × 10⁹。各指标在遗传相似性评价中表现出较高的判别性能。结论：本研究利用10个引物，基于ISSR-PCR结果，建立了未知大麻样本间遗传相似性估计的数值标准。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Forensic Toxicology TOXICOLOGY-

CiteScore

5.80

自引率

9.10%

发文量

审稿时长

3 months

期刊介绍： The journal Forensic Toxicology provides an international forum for publication of studies on toxic substances, drugs of abuse, doping agents, chemical warfare agents, and their metabolisms and analyses, which are related to laws and ethics. It includes original articles, reviews, mini-reviews, short communications, and case reports. Although a major focus of the journal is on the development or improvement of analytical methods for the above-mentioned chemicals in human matrices, appropriate studies with animal experiments are also published. Forensic Toxicology is the official publication of the Japanese Association of Forensic Toxicology (JAFT) and is the continuation of the Japanese Journal of Forensic Toxicology (ISSN 0915-9606).