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Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay ,
Author(s) -
Elkins Kelly M.,
Perez Anjelica C. U.,
Quinn Alicia A.
Publication year - 2017
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/1556-4029.13321
Subject(s) - multiplex , identification (biology) , high resolution , resolution (logic) , high resolution melt , multiplex polymerase chain reaction , chromatography , microbiology and biotechnology , chemistry , polymerase chain reaction , analytical chemistry (journal) , biology , botany , biochemistry , computer science , genetics , gene , remote sensing , geography , artificial intelligence
The international prevalence of “legal high” drugs necessitates the development of a method for their detection and identification. Herein, we describe the development and validation of a tetraplex multiplex real‐time polymerase chain reaction ( PCR ) assay used to simultaneously identify morning glory, jimson weed, Hawaiian woodrose, and marijuana detected by high‐resolution melt using LCG reen Plus ® . The PCR assay was evaluated based on the following: (i) specificity and selectivity—primers were tested on DNA extracted from 30 species and simulated forensic samples, (ii) sensitivity—serial dilutions of the target DNA were prepared, and (iii) reproducibility and reliability—sample replicates were tested and remelted on different days. The assay is ideal for cases in which inexpensive assays are needed to quickly detect and identify trace biological material present on drug paraphernalia that is too compromised for botanical microscopic identification and for which analysts are unfamiliar with the morphology of the emerging “legal high” species.