PCR-reverse dot blot of the nucleotide signature sequences of mat K for the identification of Mitragyna speciosa , a narcotic species

Jaipaew, Jirayut; Padungchareon, Thatree; Sukrong, Suchada

doi:10.1016/j.plgene.2018.04.005

Cited by 14 publications

(7 citation statements)

References 36 publications

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“…Uncorrected p-distances were calculated in Geneious, a bioinformatics desktop software package (https :// www.bioma tters .com) for sequences obtained from the trnH-psbA plastid region, which is considered as one of the most variable regions of angiosperm plants [47][48][49][50] . Comparisons were made using published sequence data from NCBI GenBank for Mitragyna spp., including M. speciosa and the newly obtained sequences in this study (Supplementary Table S5).…”

Section: Identification and Quantitative Analysis Of Alkaloids In Kramentioning

confidence: 99%

Chemical composition and biological effects of kratom (Mitragyna speciosa): In vitro studies with implications for efficacy and drug interactions

Todd

Kellogg

Wallace

et al. 2020

Sci Rep

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The safety and efficacy of kratom (Mitragyna speciosa) for treatment of pain is highly controversial. Kratom produces more than 40 structurally related alkaloids, but most studies have focused on just two of these, mitragynine and 7-hydroxymitragynine. Here, we profiled 53 commercial kratom products using untargeted LC–MS metabolomics, revealing two distinct chemotypes that contain different levels of the alkaloid speciofoline. Both chemotypes were confirmed with DNA barcoding to be M. speciosa. To evaluate the biological relevance of variable speciofoline levels in kratom, we compared the opioid receptor binding activity of speciofoline, mitragynine, and 7-hydroxymitragynine. Mitragynine and 7-hydroxymitragynine function as partial agonists of the human µ-opioid receptor, while speciofoline does not exhibit measurable binding affinity at the µ-, δ- or ƙ-opioid receptors. Importantly, mitragynine and 7-hydroxymitragynine demonstrate functional selectivity for G-protein signaling, with no measurable recruitment of β-arrestin. Overall, the study demonstrates the unique binding and functional profiles of the kratom alkaloids, suggesting potential utility for managing pain, but further studies are needed to follow up on these in vitro findings. All three kratom alkaloids tested inhibited select cytochrome P450 enzymes, suggesting a potential risk for adverse interactions when kratom is co-consumed with drugs metabolized by these enzymes.

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Section: Identification and Quantitative Analysis Of Alkaloids In Kramentioning

confidence: 99%

Chemical composition and biological effects of kratom (Mitragyna speciosa): In vitro studies with implications for efficacy and drug interactions

Todd

Kellogg

Wallace

et al. 2020

Sci Rep

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“…Previously published primers that targeted the nuclear ITS region [35] and two chloroplast barcode regions, trnH-psbA [36] and matK [34],…”

Section: Annealing Temperature Determinationmentioning

confidence: 99%

“…These curves also vary based on the target gene of interest. Lastly, a PCR-reverse dot blot assay was developed that targeted an insertion-deletion (INDEL) in the matK region of the chloroplast genome for accurate discrimination of M. speciosa from other Mitragyna species [34]. However, this method is time-consuming and laborious.…”

Section: Introductionmentioning

confidence: 99%

Sequence variation of commercially available kratom products at universal DNA barcode regions

Graham,

Cantu,

Houston

2024

Journal of Forensic Sciences

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Mitragyna speciosa, commonly known as kratom, is a narcotic plant that is used for its unique mood‐enhancing and pain‐relieving effects. It is marketed throughout the United States as a ‘legal high’ and has gained popularity as an alternative to opioids. However, kratom's increasing involvement in accidental overdoses, especially among polydrug users, has prompted warnings from the Drug Enforcement Agency (DEA) and the Food and Drug Administration (FDA). Despite these warnings, kratom remains legal federally, although it is banned in six states. This legal disparity complicates monitoring and enforcement efforts in states where kratom is illegal. Common forensic techniques using morphology or chemical analysis are beneficial in some instances but are not useful in source attribution because most seized kratom is powdered and the alkaloid content of samples can vary within products, making sourcing unreliable. This study focused on developing a DNA barcoding method to access sequence variation in commercial kratom products. It evaluated the utility of one nuclear barcode region (ITS) and three chloroplast barcode regions (matK, rbcL, and trnH‐psbA) in assessing sequence variation across commercially available kratom products. Novel polymorphisms were discovered, and the ITS region showed the greatest variation between samples. Among the 15 kratom products tested, only two haplotypes were identified across the four barcoding regions. The findings highlight the potential of DNA barcoding as a forensic tool in the traceability and enforcement against illegal kratom distribution. Nonetheless, the limited haplotypic diversity points to a need for further development and expansion of the M. speciosa DNA sequence database.

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“…However, while DNA barcoding is proven to be useful for differentiating between edible and toxic plant species (Cornara et al., 2018), there are still some associated limitations, such as being relatively time‐consuming and costly due to the need for sequencing equipment, as well as having limited or no application to plant mixtures and processed materials containing degraded DNA. Recently, DNA barcoding has been coupled with other techniques to overcome these limitations, such as reverse dot blot (Jaipaew et al., 2018), next‐generation sequencing (Dobrovolny et al., 2019), loop‐mediated isothermal amplification (Lee & Hxiao, 2019), and high‐resolution melting (HRM) analysis (Lagiotis et al., 2020; Osathanunkul et al., 2015).…”

Section: Introductionmentioning

confidence: 99%

Barcoding High Resolution Melting (Bar‐HRM) enables the discrimination between toxic plants and edible vegetables prior to consumption and after digestion

Anthoons

Lagiotis

Drouzas

et al. 2022

Journal of Food Science

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The consumption of poisonous plants can lead to serious health problems or even casualties due to various factors, including easy access to poisonous plants due to their common distribution, co-occurrence and resemblance with edible plants, and the lack of regulation in the food product supply chain. Clinical diagnosis of intoxications usually relies on the availability of the plant consumed by the patient and on the morphology of the plant parts found in the patient's stomach. Therefore, given the fragmented nature of ingested plant material, species identification may face serious difficulties, can be inaccurate, and timeconsuming. This highlights the need for rapid and reliable tools to identify toxic species. In the present study, we developed an ITS2-high-resolution melting (HRM) assay for: (1) the discrimination of common toxic plants and their edible lookalikes, and (2) the detection of toxic plants in digested samples. More specifically, we designed species-specific ITS2 primers for the authentication of poisonous species in simulated mixtures and verified them with Bar-HRM.Moreover, the developed HRM-based molecular tool was capable of quantifying the toxic species Datura stramonium in simulated mixtures with the edible Amaranthus retroflexus down to at least 0.5% v/v. This study shows that speciesspecific ITS2 primers can amplify the DNA from fragmented and/or artificially digested samples and that Bar-HRM is capable of detecting poisonous plant species in digested samples even after 4 h. The developed Bar-HRM protocol has important implications for application in medicine, forensics, and the agricultural industry, either to accurately detect the cause of plant intoxications or as a tool for quality control in the supply chain.

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PCR-reverse dot blot of the nucleotide signature sequences of mat K for the identification of Mitragyna speciosa , a narcotic species

Cited by 14 publications

References 36 publications

Chemical composition and biological effects of kratom (Mitragyna speciosa): In vitro studies with implications for efficacy and drug interactions

Chemical composition and biological effects of kratom (Mitragyna speciosa): In vitro studies with implications for efficacy and drug interactions

Sequence variation of commercially available kratom products at universal DNA barcode regions

Barcoding High Resolution Melting (Bar‐HRM) enables the discrimination between toxic plants and edible vegetables prior to consumption and after digestion

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