Genus identification of toxic plant by real-time PCR

Matsuyama, S.; Nishi, Katsuji

doi:10.1007/s00414-010-0487-8

Cited by 17 publications

(6 citation statements)

References 31 publications

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“…The DNA templates at 10 ng, 1 ng, 100 pg, 10 pg, and 1 pg showed successful amplification (6.8 ± 0.2, 8.0 ± 0.3, 8.9 ± 0.2, 10.3 ± 0.4, and 11.9 ± 0.3 min); however, amplification was not detected using 100 fg of DNA template. The sensitivity of the LAMP assay was higher than the previously reported sensitivity of real-time PCR (100 pg) [14]. We also confirmed that amplification of no-template controls did not occur for 20 min.…”

Section: Resultssupporting

confidence: 83%

“…Toxic plants can be identified by morphological analysis or chemical analysis using high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC/MS) [6, 9–11, 13]. Recently, DNA-based analytical methods, such as real time polymerase chain reaction (PCR), have been proposed for the detection of toxic plants, such as Aconitum plants and Veratrum album [14, 15]. However, these analytical methods require long processing time and a well-equipped laboratory.…”

Section: Introductionmentioning

confidence: 99%

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Rapid identification of Aconitum plants based on loop-mediated isothermal amplification assay

et al. 2019

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Objective Aconitum plants (Ranunculaceae) exhibit toxicity, and accidental ingestion of the plants has been reported in Japan. Identifying the cause of poisoning is important for emergency medical treatment, and a rapid and simple detection technique is required for the identification of poisoning cause. In the present study, we developed a rapid and simple method for detecting Aconitum plant DNA using a loop-mediated isothermal amplification (LAMP) assay. Results Specific LAMP primers for Aconitum plants were designed based on the trnL – trnF intergenic spacer region. Using the LAMP primers, the LAMP assay included an initiation reaction of 10 min followed by amplification for 20 min at the isothermal reaction temperature of 65 °C. The LAMP reaction was demonstrated to be specific and highly sensitive to Aconitum plants, given that the assay can be used for 1 pg of purified DNA. Using raw extracted DNA as template, the entire detection procedure from DNA extraction to final detection required only 30 min. Moreover, the protocol identified samples containing approximately 5 mg of Aconitum plants cooked and digested with artificial gastric juice. The currently proposed protocol exhibits good potential as a screening method of Aconitum plant poisoning for emergency medical care. Electronic supplementary material The online version of this article (10.1186/s13104-019-4463-1) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Rapid identification of Aconitum plants based on loop-mediated isothermal amplification assay

et al. 2019

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“…The artificial gastric juice was made by dissolving 2.0 g of NaCl and 3.2 g of porcine pepsin in 7.0 mL of 0.2 mol L –1 HCl. Water was added to make a total volume up to 1 L with pH of 1.2 as described by Matsuyama and Nish 21 . To make an imitation peristaltic movement, each of the grinded leaves (50 mg) was digested in 100 mL of artificial gastric juice at a 37°C in a shaking incubator at 50 r.p.m.…”

Section: Methodsmentioning

confidence: 99%

Development of molecular markers to distinguish between morphologically similar edible plants and poisonous plants using a real‐time PCR assay

Kim

Jang

2020

J Sci Food Agric

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BACKGROUNDAs a result of similar appearances between edible and poisonous plants, 42 patients have ingested poisonous plants from 2013 to 2017 in Korea. We have developed species‐specific primer sets of three of edible and poisonous plants sets (Ligularia fischeri & Caltha palustris, Artemisia annua & Ambrosia artemisiifolia and Hemerocallis fulva & Veratrum maackii) for distinguishing both plants using a real‐time polymerase chain reaction assay.RESULTSThe efficiencies of the developed primer sets ranged from 87.8% to 102.0%. The developed primer sets have significant correlation coefficient values between the Ct values and the log DNA concentration for their target species (r2 > 0.99). The cut‐off lines as the crossing point values of the limit of quantitation of the target species were determined, and all non‐target species were amplified later than the cut‐off cycles. Then, the effectiveness of the developed primer sets was evaluated using commercial food products and digested samples with simulated gastric juice.CONCLUSIONAll of the developed species‐specific primer sets were able to detect target DNA successfully in commercial food products and the digested samples. Therefore, the developed species‐specific primer sets in the present study would be useful tools for distinguishing between poisonous plants and edible plants. © 2020 Society of Chemical Industry

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“…However, there are still issues to be considered in using DNA barcoding as a clinical diagnostic tool for rapid identification of toxic plants. (1) A study has shown that the recovery of deoxyribonucleic acid from digested target plants is lower than that of untreated samples (Matsuyama and Nishi, 2011). In our study, some samples showed difficulties in amplifying the matK region after SGF digestion.…”

Section: Prospects and Problems Of Dna Barcoding As A Useful Tool For Clinical Identificationmentioning

confidence: 54%

Rapid Identification of Common Poisonous Plants in China Using DNA Barcodes

Wang

Zhao

et al. 2021

Front. Ecol. Evol.

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Toxic plants have been a major threat to public health in China. However, identification and tracing of poisoned species with traditional methods are unreliable due to the destruction of plant morphology by cooking and chewing. DNA barcoding is independent of environmental factors and morphological limitations, making it a powerful tool to accurately identify species. In our study, a total of 83 materials from 26 genera and 31 species of 13 families were collected and 13 plant materials were subjected to simulated gastric fluid digestion. Four markers (rbcL, trnH-psbA, matK, and ITS) were amplified and sequenced for all untreated and mock-digested samples. The effectiveness of DNA barcoding for the identification of toxic plants was assessed using Basic Local Alignment Search Tool (BLAST) method, PWG-Distance method, and Tree-Building (NJ) method. Except for the matK region, the amplification success rate of the remaining three regions was high, but the sequencing of trnH-psbA and ITS was less satisfactory. Meanwhile, matK was prone to be more difficult to amplify and sequence because of simulated gastric fluid. Among the three methods applied, BLAST method showed lower recognition rates, while PWG-Distance and Tree-Building methods showed little difference in recognition rates. Overall, ITS had the highest recognition rate among individual loci. Among the combined loci, rbcL + ITS had the highest species recognition rate. However, the ITS region may not be suitable for DNA analysis of gastric contents and the combination of loci does not significantly improve species resolution. In addition, identification of species to the genus level is sufficient to aid in the clinical management of most poisoning events. Considering primer versatility, DNA sequence quality, species identification ability, experimental cost and speed of analysis, we recommend rbcL as the best single marker for clinical identification and also suggest the BLAST method for analysis. Our current results suggest that DNA barcoding can rapidly identify and trace toxic species and has great potential for clinical applications. In addition, we suggest the creation of a proprietary database containing morphological, toxicological and molecular information to better apply DNA barcoding technology in clinical diagnostics.

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Genus identification of toxic plant by real-time PCR

Cited by 17 publications

References 31 publications

Rapid identification of Aconitum plants based on loop-mediated isothermal amplification assay

Rapid identification of Aconitum plants based on loop-mediated isothermal amplification assay

Development of molecular markers to distinguish between morphologically similar edible plants and poisonous plants using a real‐time PCR assay

Rapid Identification of Common Poisonous Plants in China Using DNA Barcodes

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