Evaluation of DNA barcoding coupled high resolution melting for discrimination of closely related species in phytopharmaceuticals

Osathanunkul, Maslin; Suwannapoom, Chatmongkon; Osathanunkul, Kitisak; Madesis, Panagiotis; Boer, Hugo de

doi:10.1016/j.phymed.2015.11.018

Cited by 47 publications

(27 citation statements)

References 48 publications

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“…HRM technology has been utilized for discriminating cultivated varieties, medicinal plant species, herbal tea, wine authenticity (Jung et al, 2010; Osathanunkul et al, 2015, 2016; Costa et al, 2016; Li et al, 2016; Song et al, 2016; Pereira et al, 2017). However, a reliable HRM approach to identify herbal medicines on Chinese herb market is still lacks.…”

Section: Resultsmentioning

confidence: 99%

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Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology

Sun

Yan

et al. 2017

Front. Plant Sci.

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There is an unmet need for herbal medicine identification using a fast, sensitive, and easy-to-use method that does not require complex infrastructure and well-trained technicians. For instance, the detection of adulterants in Lobelia chinensis herbal product has been challenging, since current detection technologies are not effective due to their own limits. High Resolution Melting (HRM) has emerged as a powerful new technology for clinical diagnosis, research in the food industry and in plant molecular biology, and this method has already highlighted the complexity of species identification. In this study, we developed a method of species specific detection of L. chinensis using HRM analysis combined with internal transcribed spacer 2. We then applied this method to commercial products purporting to contain L. chinensis. Our results demonstrated that HRM can differentiate L. chinensis from six common adulterants. HRM was proven to be a fast and accurate technique for testing the authenticity of L. chinensis in herbal products. Based on these results, a HRM approach for herbal authentication is provided.

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Section: Resultsmentioning

confidence: 99%

“…However, it is difficult to utilize HRM technology to differentiate species with small differences in meting temperatures (<0.25°C) and homozygotes with C to G and A to T changes in specific site (Simko, 2016). Another single barcode or multi-locus combination can provide optimized discrimination (Osathanunkul et al, 2016). …”

Section: Resultsmentioning

confidence: 99%

Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology

Sun

Yan

et al. 2017

Front. Plant Sci.

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“…Bar-HRM was proved to be a powerful tool for species identification that is capable not only to identify but also to quantitatively detect adulterants. The power of Bar-HRM have been demonstrated in bean crops [ 18 ], Hazel nut [ 19 ], Berry [ 20 ], and recently authenticate medicinal plant species in herbal products [ 21 – 25 ]. The method has been only used to detect or identify adulteration or inferior quality in finished products.…”

Section: Introductionmentioning

confidence: 99%

Should DNA sequence be incorporated with other taxonomical data for routine identifying of plant species?

Suesatpanit

Osathanunkul

Madesis

et al. 2017

BMC Complement Altern Med

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BackgroundA variety of plants in Acanthaceae have long been used in traditional Thai ailment and commercialised with significant economic value. Nowadays medicinal plants are sold in processed forms and thus morphological authentication is almost impossible. Full identification requires comparison of the specimen with some authoritative sources, such as a full and accurate description and verification of the species deposited in herbarium. Intake of wrong herbals can cause adverse effects. Identification of both raw materials and end products is therefore needed.MethodsHere, the potential of a DNA-based identification method, called Bar-HRM (DNA barcoding coupled with High Resolution Melting analysis), in raw material species identification is investigated. DNA barcode sequences from five regions (matK, rbcL, trnH-psbA spacer region, trnL and ITS2) of Acanthaceae species were retrieved for in silico analysis. Then the specific primer pairs were used in HRM assay to generate unique melting profiles for each plants species.ResultsThe method allows identification of samples lacking necessary morphological parts. In silico analyses of all five selected regions suggested that ITS2 is the most suitable marker for Bar-HRM in this study. The HRM analysis on dried samples of 16 Acanthaceae medicinal species was then performed using primer pair derived from ITS2 region. 100% discrimination of the tested samples at both genus and species level was observed. However, two samples documented as Clinacanthus nutans and Clinacanthus siamensis were recognised as the same species from the HRM analysis. Further investigation reveals that C. siamensis is now accepted as C. nutans.ConclusionsThe results here proved that Bar-HRM is a promising technique in species identification of the studied medicinal plants in Acanthaceae. In addition, molecular biological data is currently used in plant taxonomy and increasingly popular in recent years. Here, DNA barcode sequence data should be incorporated with morphological characters in the species identification.

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“…Furthermore, DNA is more informative than chemical composition and can be easily extracted in the presence of small traces of organic materials. Presently, methods that involve molecular biological techniques, such as sequence characterized amplified region markers (Adinolfi et al, 2007;Basha et al, 2009), amplified fragment length polymorphism (Percifield et al, 2008;Bandyopadhyay and Raychaudhuri, 2012), DNA microarray (Zhang et al, 2003;Liu et al, 2012), DNA barcoding (Chen et al, 2010;Mahadani and Ghosh, 2013;Pang et al, 2013;Xin et al, 2015), and high resolution melting (Osathanunkul et al, 2016) are available for herbal medicine identification. The major advantages of these methods are rapidity, high sensitivity, and specificity.…”

Section: Introductionmentioning

confidence: 99%

Loop-Mediated Isothermal Amplification (LAMP): Emergence As an Alternative Technology for Herbal Medicine Identification

Xiong

Liu

et al. 2016

Front. Plant Sci.

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Correct identification of medicinal plant ingredients is essential for their safe use and for the regulation of herbal drug supply chain. Loop-mediated isothermal amplification (LAMP) is a recently developed approach to identify herbal medicine species. This novel molecular biology technique enables timely and accurate testing, especially in settings where infrastructures to support polymerase chain reaction facilities are lacking. Studies that used this method have altered our view on the extent and complexity of herbal medicine identification. In this review, we give an introduction into LAMP analysis, covers the basic principles and important aspects in the development of LAMP analysis method. Then we presented a critical review of the application of LAMP-based methods in detecting and identifying raw medicinal plant materials and their processed products. We also provide a practical standard operating procedure (SOP) for the utilization of the LAMP protocol in herbal authentication, and consider the prospects of LAMP technology in the future developments of herbal medicine identification and the challenges associated with its application.

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Evaluation of DNA barcoding coupled high resolution melting for discrimination of closely related species in phytopharmaceuticals

Cited by 47 publications

References 48 publications

Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology

Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology

Should DNA sequence be incorporated with other taxonomical data for routine identifying of plant species?

Loop-Mediated Isothermal Amplification (LAMP): Emergence As an Alternative Technology for Herbal Medicine Identification

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