Simultaneous Identification of Four “Legal High” Plant Species in a Multiplex PCR High‐Resolution Melt Assay^,

A tetraplex multiplex real-time polymerase chain reaction (PCR) assay was developed to identify four 'legal high' drugs simultaneously.

• The assay detects morning glory, jimson weed, Hawaiian woodrose, and marijuana using high-resolution melt technology.
• Specificity and selectivity were confirmed by testing primers on DNA from 30 different species and simulated forensic samples.
• Sensitivity was assessed using serial dilutions of the target DNA.
• Reproducibility and reliability were established through testing sample replicates over multiple days.
• The method is suitable for identifying trace biological material on drug paraphernalia when traditional identification methods are inadequate.

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