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

1
Department of Forensic Science, Medical School of Soochow University, Suzhou 215123, China
2
Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Science, Ministry of Justice, Shanghai 200063, China
3
Department of Forensic Medicine, School of Basic Medical Science, Wenzhou Medical University, Wenzhou 325035, China
4
Health Science Center, College of Medicine and Forensics, Xi’an Jiaotong University, Xi’an 710061, China
*
Authors to whom correspondence should be addressed.
Received: 7 December 2020 / Revised: 18 January 2021 / Accepted: 26 January 2021 / Published: 29 January 2021
(This article belongs to the Section Technologies and Resources for Genetics)
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. View Full-Text
Keywords: polymerase chain reaction (PCR); high-resolution melting (HRM); DNA barcoding; internal transcribed spacer (ITS); Psilocybe cubensis polymerase chain reaction (PCR); high-resolution melting (HRM); DNA barcoding; internal transcribed spacer (ITS); Psilocybe cubensis
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MDPI and ACS Style

Zhang, X.; Yu, H.; Yang, Q.; Wang, Z.; Xia, R.; Chen, C.; Qu, Y.; Tan, R.; Shi, Y.; Xiang, P.; Zhang, S.; Li, C. A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis. Genes 2021, 12, 199. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12020199

AMA Style

Zhang X, Yu H, Yang Q, Wang Z, Xia R, Chen C, Qu Y, Tan R, Shi Y, Xiang P, Zhang S, Li C. A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis. Genes. 2021; 12(2):199. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12020199

Chicago/Turabian Style

Zhang, Xiaochun, Huan Yu, Qi Yang, Ziwei Wang, Ruocheng Xia, Chong Chen, Yiling Qu, Rui Tan, Yan Shi, Ping Xiang, Suhua Zhang, and Chengtao Li. 2021. "A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis" Genes 12, no. 2: 199. https://0-doi-org.brum.beds.ac.uk/10.3390/genes12020199

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