DNA barcoding in herbal medicine: Retrospective and prospective

Chen, Shilin; Yin, Xiao; Han, Jianping; Sun, Wei; Yao, Hui; Song, Jingyuan; Li, Xiwen

doi:10.1016/j.jpha.2023.03.008

Cited by 21 publications

(9 citation statements)

References 153 publications

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“…The ITS2 sequence can identify medicinal plants and counterfeits at the species level, which is an effective method for monitoring the authenticity of traditional Chinese medicine. 44 The researchers demonstrated that ITS2 sequences serve as DNA barcodes to help identify the original plant of LJF and LF. 45 However, ITS2-based DNA barcoding has a low success rate when detecting mixed samples.…”

Section: Market Sample Inspectionsmentioning

confidence: 99%

Rapid and simultaneous identification of Lonicera Japonicae Flos and Lonicerae Flos using MLPA‐HRM and rhPCR‐HRM

Wang

et al. 2023

JSFA Reports

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Background: Lonicera Japonicae Flos (LJF) and Lonicerae Flos (LF), derived from different species of Lonicera, are two different types of traditional Chinese medicine (TCM) according to their differences in composition and effectiveness. Because of the high price, good effectiveness of the medicinal, and functional values, LJF is often adulterated by LF in the market. Therefore, two single nucleotide polymorphism-based genotyping methods, multiplex ligation-dependent probe amplification high-resolution melting (MLPA-HRM) and RNase H2-dependant PCR (rhPCR)-HRM, were established for the identification of LF and LJF.Results: Highly specific MLPA probes and rhPCR-primers were designed based on the trnL-trnF regions. The melting temperature (T m ) of amplicons was derived from the peak of the HRM curve, and the differences in T m values were used to identify LJF and LF. The two methods have both shown good specificity for identifying LJF and LF without cross-reaction, and have high sensitivity with the detection limits of 0.1 ng DNA template. For mixed samples, MLPA-HRM could detect LJF mixed with 10% LF while rhPCR-HRM could detect 5% LF in mixed samples. The MLPA-HRM and rhPCR-HRM were performed on DNA extracted from 54 samples of LJF and LF randomly selected from the medicinal material market, all samples were successfully identified and further verified with DNA barcoding. Conclusion:MLPA-HRM and rhPCR-HRM methods developed in our study not only provided fast, simple, sensitive, and practical identification of LJF and LF, but also provided technical references for the adulteration identification of other herb-medicines, they are of great significance to the quality control and safe use of plant medicinal materials.

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Section: Market Sample Inspectionsmentioning

confidence: 99%

Rapid and simultaneous identification of Lonicera Japonicae Flos and Lonicerae Flos using MLPA‐HRM and rhPCR‐HRM

Wang

et al. 2023

JSFA Reports

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“…Traditional clinical laboratory identification based on morphology is limited to the section or species-complex level for Aspergillus , making it challenging to achieve species-level identification (Gautier et al, 2016 ). DNA barcoding, particularly utilizing the ITS region, has become widely employed for fungal identification (Fernandes et al, 2021 ; Chen et al, 2023 ). However, due to limited polymorphism, secondary markers such as partial β -tubulin or calmodulin are needed for confident species identification, albeit within a narrow taxonomic range (Geiser et al, 2007 ; Schoch et al, 2012 ; Gautier et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Identification of closely related species in Aspergillus through Analysis of Whole-Genome

Qi,

Hao,

Gan

et al. 2024

Front. Microbiol.

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The challenge of discriminating closely related species persists, notably within clinical diagnostic laboratories for invasive aspergillosis (IA)-related species and food contamination microorganisms with toxin-producing potential. We employed Analysis of the whole-GEnome (AGE) to address the challenges of closely related species within the genus Aspergillus and developed a rapid detection method. First, reliable whole genome data for 77 Aspergillus species were downloaded from the database, and through bioinformatic analysis, specific targets for each species were identified. Subsequently, sequencing was employed to validate these specific targets. Additionally, we developed an on-site detection method targeting a specific target using a genome editing system. Our results indicate that AGE has successfully achieved reliable identification of all IA-related species (Aspergillus fumigatus, Aspergillus niger, Aspergillus nidulans, Aspergillus flavus, and Aspergillus terreus) and three well-known species (A. flavus, Aspergillus parasiticus, and Aspergillus oryzae) within the Aspergillus section. Flavi and AGE have provided species-level-specific targets for 77 species within the genus Aspergillus. Based on these reference targets, the sequencing results targeting specific targets substantiate the efficacy of distinguishing the focal species from its closely related species. Notably, the amalgamation of room-temperature amplification and genome editing techniques demonstrates the capacity for rapid and accurate identification of genomic DNA samples at a concentration as low as 0.1 ng/μl within a concise 30-min timeframe. Importantly, this methodology circumvents the reliance on large specialized instrumentation by presenting a singular tube operational modality and allowing for visualized result assessment. These advancements aptly meet the exigencies of on-site detection requirements for the specified species, facilitating prompt diagnosis and food quality monitoring. Moreover, as an identification method based on species-specific genomic sequences, AGE shows promising potential as an effective tool for epidemiological research and species classification.

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“…The COI gene is widely used in the identi cation of animals as a universal DNA barcode 12,13 . However, universal DNA barcoding has certain limitations: it is incapable of identifying the species of herbal materials in Chinese patent medicine (CPM) and proprietary Chinese medicinal materials that have undergone severe DNA degradation 14 . Unfortunately, the medicinal material of leeches typically undergoes drying or high temperature processing during preparation, which leads to DNA degradation.…”

Section: Introductionmentioning

confidence: 99%

Advancing Species Identification in Traditional Chinese Medicine: A Novel Mini-Barcode from Mitochondrial Genomes Combined with Meta-barcoding Technology for Leech Authentication

Shi,

Guo,

Yao

et al. 2024

Preprint

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In Traditional Chinese Medicine (TCM), the medicinal leech is a vital component for treatments aimed at promoting blood circulation and eliminating blood stasis. However, the prevalence of counterfeit leech products in the market undermines the quality and efficacy of these remedies. Conventional DNA barcoding techniques, such as the COI barcode, have proven inadequate for TCM species identification due to amplification challenges. This study introduces a novel 219bp mini-barcode, derived from the mitochondrial genome's 16S rRNA gene, exhibiting high variability among five leech species. We also used primers 741F-943R to enhance the identification process. Compared with the traditional COI barcode, our mini-barcode showed remarkable identification efficiency, accurately classifying 144 out of 147 leech samples from fresh and processed materials. In the case of seven batches of leech decoction pieces, the mini-barcode identified six, whereas the COI barcode only recognized one. Additionally, when combined with meta-barcoding technology, the mini-barcode effectively discerned five leech species within Chinese patent medicines. These results confirm the mini-barcode's potential as a reliable tool for the rapid and precise identification of leech species in TCM products.

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DNA barcoding in herbal medicine: Retrospective and prospective

Cited by 21 publications

References 153 publications

Rapid and simultaneous identification of Lonicera Japonicae Flos and Lonicerae Flos using MLPA‐HRM and rhPCR‐HRM

Rapid and simultaneous identification of Lonicera Japonicae Flos and Lonicerae Flos using MLPA‐HRM and rhPCR‐HRM

Identification of closely related species in Aspergillus through Analysis of Whole-Genome

Advancing Species Identification in Traditional Chinese Medicine: A Novel Mini-Barcode from Mitochondrial Genomes Combined with Meta-barcoding Technology for Leech Authentication

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