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

J Agric Food Chem. 2013 Feb 6;61(5):1060-6. doi: 10.1021/jf304223a. Epub 2013 Jan 23.

Abstract

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) format to detect a single nucleotide polymorphism (SNP) unique to this species. This assay was able to detect P. armandii in homogenates down 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.

MeSH terms

  • DNA, Plant / isolation & purification
  • Dysgeusia / etiology
  • Dysgeusia / physiopathology*
  • Genome, Chloroplast / genetics
  • Humans
  • Nuts*
  • Oligonucleotides / isolation & purification*
  • Pinus / chemistry
  • Pinus / genetics*
  • Polymorphism, Single Nucleotide
  • Real-Time Polymerase Chain Reaction / methods*
  • Selection, Genetic
  • Sequence Alignment
  • Taste

Substances

  • DNA, Plant
  • Oligonucleotides
  • locked nucleic acid