Development of a Locked Nucleic Acid Real-Time Polymerase Chain Reaction Assay for the Detection of Pinus armandii in Mixed Species Pine Nut Samples Associated with Dysgeusia

Recent work has shown that the presence of the species Pinus armandii, even when occurring as species mixtures of pine nuts, is correlated with taste disturbance (dysgeusia), also referred to as "pine mouth". Because of this known possibility of pine nut mixtures, a need was identified for a rapid streamlined assay to detect the presence of this species in the presence of other types of pine nuts. A locked nucleic acid probe was employed in a real-time polymerase chain reaction (RT-PCR) forrnat to detect a single nucleotide polymorphism (SNP) unique to this species. This assay was able to detect P. armandii in homogenates down to similar to 1% concentration (the lowest level tested) in the presence of several commonly co-occurring and closely related species of pine and should prove to be a useful tool for the detection of this species in food products.