| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Outline of Final Research Achievements |
Veratrum album is one of the most common causative plants of poisoning by accidental ingestion in Japan. In this study, rapid and specific detection methods for identification of V. album by real-time PCR and Loop-Mediated Isothermal Amplification (LAMP) were developed.
First, direct sequencing of specific loci for plant species identification, known as DNA barcoding to search for species-specific regions was performed, and the results indicated that trnH-psbA and trnL-trnF regions were useful for identification of V. album.I next developed real-time PCR to detect V. album. 2 primer sets utilizing species-specific sequences in trnH-psbA and trnL-trnF were designed. One primer set, utilizing trnH-psbA, could detect 1 pg V. album DNA, and another primer set, utilizing trnL-trnF, could detect 10 pg DNA. Both primer sets were specific for V. album. Finally, the LAMP primer set for trnL-trnF region was designed. The detection limit was 80 pg, and the primer set was specific for V. album.
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