Rapid and sensitive detection of Lophophora williamsii by loop-mediated isothermal amplification.
We have developed a convenient method for the detection of Lophophora williamsii using loop-mediated isothermal amplification (LAMP). We designed six species-specific primers for L. williamsii, including two loop primers. This L. williamsii-specific primer set was used for LAMP of total DNA extracted from L. williamsii and from L. diffusa. Real-time monitoring of LAMP was achieved by measuring turbidity due to the formation of magnesium pyrophosphate. Amplification occurred in samples mixed with total DNA from L. williamsii, but not in those mixed with total DNA from L. diffusa. We could also visually observe L. williamsii by adding fluorescent detection reagent to the reaction tube and exposing it to UV light. This new method amplified L. williamsii selectively and is expected to be applicable to the detection of peyote.
Department of Medicinal Plant Science, Hoshi University, Shinagawa, Tokyo, Japan.
Yohei
Y
Sasaki
Tsuguto
T
Fujimoto
Masako
M
Aragane
Ichiro
I
Yasuda
Seiji
S
Nagumo
D001483:Base Sequence; D029421:Cactaceae / Q000145:classification* / Q000235:genetics; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009391:Nephelometry and Turbidimetry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D018744:DNA, Plant