title: Simultaneous Identification of Four "Legal High" Plant Species in a Multiplex PCR High-Resolution Melt Assay.
abstract: 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 LCGreen 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.
authors.affiliation: Chemistry Department, Forensic Science Program, Towson University, 8000 York Road, Towson, MD, 21252.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Elkins
authors.affiliation: Chemistry Department, Forensic Science Program, Towson University, 8000 York Road, Towson, MD, 21252.
authors.forename: Anjelica C U
authors.initials: ACU
authors.lastname: Perez
authors.affiliation: Chemistry Department, Forensic Science Program, Towson University, 8000 York Road, Towson, MD, 21252.
authors.forename: Alicia A
authors.initials: AA
authors.lastname: Quinn
mesh_terms: D002188:Cannabis / Q000235:genetics*; D027722:Convolvulaceae / Q000235:genetics*; D017931:DNA Primers; D013286:Datura stramonium / Q000235:genetics*; D044707:Forensic Sciences; D006801:Humans; D027741:Ipomoea / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019966:Substance-Related Disorders; D013696:Temperature
chemical_list: D017931:DNA Primers
keywords: 
  Cannabis sativa
  ; 
  Datura stramonium
  ; 
  Ipomoea purpurea
  ; 
  Merremia tuberosa
  ; LCGreen Plus®; SYBR green I; forensic science; high-resolution melt; polymerase chain reaction