Development of Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Cannabis sativa

被引：19
作者：
Kitamura, Masashi [1 ,2 ]
Aragane, Masako [3 ]
Nakamura, Kou [3 ]
Watanabe, Kazuhito [4 ]
Sasaki, Yohei [1 ]
机构：
[1] Kanazawa Univ, Grad Sch Med Sci, Div Pharmaceut Sci, Lab Mol Pharmacognosy, Kakuma Machi, Kanazawa, Ishikawa 9201192, Japan
[2] Ishikawa Prefectural Police HQ, Forens Sci Lab, 1-1 Kuratsuki, Kanazawa, Ishikawa 9208553, Japan
[3] Tokyo Metropolitan Inst Publ Hlth, Med Plant Garden, 21-1 Nakajima Cho, Tokyo 1870033, Japan
[4] Hokuriku Univ, Fac Pharmaceut Sci, Ho 3 Kanagawa Machi, Kanazawa, Ishikawa 9201181, Japan
来源：
BIOLOGICAL & PHARMACEUTICAL BULLETIN | 2016年 / 39卷 / 07期
关键词：
Cannabis sativa; isothermal amplification; tetrahydrocannabinolic acid; loop-mediated isothermal amplification; marijuana; hemp; IDENTIFICATION; DNA; MARIJUANA; VIRUS; CONSTITUENTS; EXTRACTS; SYNTHASE; PLANTS; GENE; DRUG;
D O I：
10.1248/bpb.b16-00090
中图分类号：
R9 [药学];
学科分类号：
1007 ;
摘要：
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 degrees 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.
引用
收藏
页码：1144 / 1149
页数：6
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