Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Cannabis sativa.
In many parts of the world, the possession and cultivation of Cannabis sativa L. are restricted by law. As chemical or morphological analyses cannot identify the plant in some cases, a simple yet accurate DNA-based method for identifying C. sativa is desired. We have developed a loop-mediated isothermal amplification (LAMP) assay for the rapid identification of C. sativa. By optimizing the conditions for the LAMP reaction that targets a highly conserved region of tetrahydrocannabinolic acid (THCA) synthase gene, C. sativa was identified within 50 min at 60-66°C. The detection limit was the same as or higher than that of conventional PCR. The LAMP assay detected all 21 specimens of C. sativa, showing high specificity. Using a simple protocol, the identification of C. sativa could be accomplished within 90 min from sample treatment to detection without use of special equipment. A rapid, sensitive, highly specific, and convenient method for detecting and identifying C. sativa has been developed and is applicable to forensic investigations and industrial quality control.
Laboratory of Molecular Pharmacognosy, Division of Pharmaceutical Sciences, Graduate School of Medical Sciences, Kanazawa University.
Masashi
M
Kitamura
Masako
M
Aragane
Kou
K
Nakamura
Kazuhito
K
Watanabe
Yohei
Y
Sasaki
D002188:Cannabis / Q000235:genetics*; D018744:DNA, Plant / Q000032:analysis; D019746:Intramolecular Oxidoreductases / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D018515:Plant Leaves / Q000235:genetics
D018744:DNA, Plant; D019746:Intramolecular Oxidoreductases; C100677:delta(1)-tetrahydrocannabinolic acid synthase