Psychedelic fungus (Psilocybe sp.) authentication in a case of illegal drug traffic: sporological, molecular analysis and identification of the psychoactive substance

Solano, Jaime; Anabalón, Leonardo; Figueroa, Sylvia; Lizama, Cristián; Reyes, Luis Chávez; Gangitano, David

doi:10.1016/j.scijus.2018.08.005

Cited by 12 publications

(10 citation statements)

References 11 publications

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“…The HRM melting curve has prominent capacity to discriminate DNA barcodes based on their composition, length, or GC content, which allows for the examination of genetic variations within DNA sequences. Moreover, it supports forensic evidence of the suspicious species [19][20][21][22][23]. For example, it has been demonstrated that HRM could identify Cannabis sativa and "legal high" drugs, among other species [21].…”

Section: Introductionsupporting

confidence: 62%

“…The identification of fungi at the species level is critical for many research areas, such as health sciences, agriculture, and forensic science, and for determining whether hallucinogenic mushrooms are essential for treatment or are being trafficked as drugs [13,16,22]. In addition, studies have shown that psilocybin may relieve depression and so this HRM method could also be utilized for research in this area [30].…”

Section: Discussionmentioning

confidence: 99%

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A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Zhang

Yang

et al. 2021

Genes

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In recent years, trafficking and abuse of hallucinogenic mushrooms have become a serious social problem. It is therefore imperative to identify hallucinogenic mushrooms of the genus Psilocybe for national drug control legislation. An internal transcribed spacer (ITS) is a DNA barcoding tool utilized for species identification. Many methods have been used to discriminate the ITS region, but they are often limited by having a low resolution. In this study, we sought to analyze the ITS and its fragments, ITS1 and ITS2, by using high-resolution melting (HRM) analysis, which is a rapid and sensitive method for evaluating sequence variation within PCR amplicons. The ITS HRM assay was tested for specificity, reproducibility, sensitivity, and the capacity to analyze mixture samples. It was shown that the melting temperatures of the ITS, ITS1, and ITS2 of Psilocybe cubensis were 83.72 ± 0.01, 80.98 ± 0.06, and 83.46 ± 0.08 °C, and for other species, we also obtained species-specific results. Finally, we performed ITS sequencing to validate the presumptive taxonomic identity of our samples, and the sequencing output significantly supported our HRM data. Taken together, these results indicate that the HRM method can quickly distinguish the DNA barcoding of Psilocybe cubensis and other fungi, which can be utilized for drug trafficking cases and forensic science.

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Section: Introductionsupporting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Zhang

Yang

et al. 2021

Genes

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“…The reaction mixture for real‐time PCR and HRM analysis consisted of a total volume of 15 µl. This contained 7.5 µl of 2× Kapa SYBR qPCR Mix (Kapa Biosystems Roche), (Cowan & Elkins, ; Solano et al, ) 0.2 µM forward primer, 0.2 µM reverse primer, and 1 µl of 25 ng/µl DNA. SYBR fluorescence dye was used to monitor the accumulation of the amplified product during PCR and the HRM process to derive the T m value.…”

Section: Methodsmentioning

confidence: 99%

High‐resolution melting of the cytochrome B gene in fecal DNA: A powerful approach for fox species identification of the Lycalopex genus in Chile

Anabalón

Encina‐Montoya

Sánchez

et al. 2019

Ecology and Evolution

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Easy, economic, precise species authentication is currently necessary in many areas of research and diagnosis in molecular biology applied to conservation studies of endangered species. Here, we present a new method for the identification of three fox species of the Lycalopex genus in Chile. We developed an assay based on high‐resolution melt analysis of the mitochondrial cytochrome B gene, allowing a simple, low cost, fast, and accurate species determination. To validate the assay applicability for noninvasive samples, we collected fecal samples in the Atacama Desert, finding unexpectedly one species outside of its known distribution range. We conclude that the assay has a potential to become a valuable tool for a standardized genetic monitoring of the Lycalopex species in Chile.

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“…HRM analysis is the method of choice for rapid analysis of sequence variation within PCR amplicons by determining their melting temperature (T m ). 22 T m depends on the base composition of amplicons. T m differences can be detected by monitoring the fluorescence changes during PCR cycles using real-time PCR differential analysis.…”

Section: Introductionmentioning

confidence: 99%

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

Anabalón

Solano

Encina‐Montoya

et al. 2022

Cannabis and Cannabinoid Research

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Background: Cannabis plants and their seed have been used in many cultures as a source of medicine and feeding during history. Today, there is an increasing demand for cannabis seeds for medical use. Moreover, a seed sales market with no legal regulations has also grown. This may pose some issues if a quality control is not set in place. Identification of cannabis strains is important for quality control purposes in a nonregulated growing market and in cases of illegal traffic and medical use. Owing to the high price as a pharmacological drug, commercial products of cannabis plants and seeds for medical users are often subjected to adulterations, either when packing or distributing certified seeds in the market. Materials and Methods: Cannabis commercial seeds and cannabis seeds for medical use were analyzed with high-resolution melting (HRM) analysis using barcoding markers. Humulus lupulus L. plants from a local market were used as outgroup control. DNA barcoding uses specific regions of the genome to identify differences in the genetic sequence of conserved regions such as internal transcribed spacer (ITS) and rbcL. DNA barcoding data can be generated with real-time polymerase chain reaction combined with HRM analysis to distinguish specific conserved DNA regions of closely related species. HRM analysis is the method of choice for rapid analysis of sequence variation. Results: The melting temperature (T m ) of homogeneous packages was consistent with single genotypes. However, packages containing contaminating seeds showed T m differences of 0.2°C on average. Conclusions: An effective, rapid, and low-cost method based on ITS nuclear DNA and on chloroplast rbcL regions for screening and detection of contamination in commercial cannabis seeds was developed and applied for the analysis of different samples. This approach can be used as a quality control tool for cannabis seeds or other plant material.

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Psychedelic fungus (Psilocybe sp.) authentication in a case of illegal drug traffic: sporological, molecular analysis and identification of the psychoactive substance

Cited by 12 publications

References 11 publications

A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

High‐resolution melting of the cytochrome B gene in fecal DNA: A powerful approach for fox species identification of the Lycalopex genus in Chile

Cannabis Seeds Authentication by Chloroplast and Nuclear DNA Analysis Coupled with High-Resolution Melting Method for Quality Control Purposes

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