Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea
Adulteration by Bacopa monnieri (BM) in Portulaca oleracea (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in t...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-02-01
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| Series: | Molecular and Cellular Probes |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0890850822001013 |
| _version_ | 1797788459185733632 |
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| author | Mo-Rong Xu Meng-Shiou Lee Bo-Cheng Yang Hsiu-Chi Chang Chao-Lin Kuo Chia-Hsin Lin Hsi-Jien Chen Jai-Hong Cheng Fang-Chun Sun |
| author_facet | Mo-Rong Xu Meng-Shiou Lee Bo-Cheng Yang Hsiu-Chi Chang Chao-Lin Kuo Chia-Hsin Lin Hsi-Jien Chen Jai-Hong Cheng Fang-Chun Sun |
| author_sort | Mo-Rong Xu |
| collection | DOAJ |
| description | Adulteration by Bacopa monnieri (BM) in Portulaca oleracea (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in the herbal market. The sequence divergences in internal transcribed spacer 2 (ITS2) between PO and its adulterant species were used to design diagnostic PCR primers. The specific designed primer sets were evaluated and show that the diagnostic PCR assay can be used to verify the authenticity of PO and BM. The detection limits of the primer set for PO and BM identification were 10 pg and 1 pg, respectively. The reactivity of diagnostic PCR was 0.1% PO genomic DNA and 0.01% BM genomic DNA in the test sample during DNA amplification. In addition, multiplex PCR (mPCR) for PO and BM identification was also established. The samples were more susceptible to the effect of steaming in authentication by singleplex PCR and mPCR than boiling and drying treatment. Furthermore, commercial samples from the market were used to demonstrate the applicability of the developed diagnostic PCR for PO authentication and diagnose BM adulteration, and the investigation found that approximately 72.2% (13/18) of PO plants in the herbal market were adulterated. In conclusion, the diagnostic PCR assay was successfully developed and its specificity, sensitivity and reactivity for PO and BM authentication were proven. These developed PCR-based molecular methods can be applied as an identification tool for PO authenticity and can be practically applied for inspection of BM adulteration in the herbal market in the future. |
| first_indexed | 2024-03-13T01:36:51Z |
| format | Article |
| id | doaj.art-625a2794d15c434f9c5024ae9ddcc57d |
| institution | Directory Open Access Journal |
| issn | 0890-8508 |
| language | English |
| last_indexed | 2024-03-13T01:36:51Z |
| publishDate | 2023-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular and Cellular Probes |
| spelling | doaj.art-625a2794d15c434f9c5024ae9ddcc57d2023-07-04T05:09:07ZengElsevierMolecular and Cellular Probes0890-85082023-02-0167101890Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleraceaMo-Rong Xu0Meng-Shiou Lee1Bo-Cheng Yang2Hsiu-Chi Chang3Chao-Lin Kuo4Chia-Hsin Lin5Hsi-Jien Chen6Jai-Hong Cheng7Fang-Chun Sun8China Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanChina Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanChina Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanChina Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanChina Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanChina Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, TaiwanDepartment of Safety, Health, and Environmental Engineering, Ming Chi University of Technology, New Taipei City, 24301, TaiwanCenter for Shockwave Medicine and Tissue Engineering, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, 833, TaiwanDepartment of Bioresources, Department of Medicinal Botanicals and Foods on Health Applicaitons, Da-Yeh University, Changhua, 515006, Taiwan; Corresponding author.Adulteration by Bacopa monnieri (BM) in Portulaca oleracea (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in the herbal market. The sequence divergences in internal transcribed spacer 2 (ITS2) between PO and its adulterant species were used to design diagnostic PCR primers. The specific designed primer sets were evaluated and show that the diagnostic PCR assay can be used to verify the authenticity of PO and BM. The detection limits of the primer set for PO and BM identification were 10 pg and 1 pg, respectively. The reactivity of diagnostic PCR was 0.1% PO genomic DNA and 0.01% BM genomic DNA in the test sample during DNA amplification. In addition, multiplex PCR (mPCR) for PO and BM identification was also established. The samples were more susceptible to the effect of steaming in authentication by singleplex PCR and mPCR than boiling and drying treatment. Furthermore, commercial samples from the market were used to demonstrate the applicability of the developed diagnostic PCR for PO authentication and diagnose BM adulteration, and the investigation found that approximately 72.2% (13/18) of PO plants in the herbal market were adulterated. In conclusion, the diagnostic PCR assay was successfully developed and its specificity, sensitivity and reactivity for PO and BM authentication were proven. These developed PCR-based molecular methods can be applied as an identification tool for PO authenticity and can be practically applied for inspection of BM adulteration in the herbal market in the future.http://www.sciencedirect.com/science/article/pii/S0890850822001013Portulaca oleraceaBacopa monnieriDiagnostic PCRTraditional Chinese medicine (TCM)Authentication |
| spellingShingle | Mo-Rong Xu Meng-Shiou Lee Bo-Cheng Yang Hsiu-Chi Chang Chao-Lin Kuo Chia-Hsin Lin Hsi-Jien Chen Jai-Hong Cheng Fang-Chun Sun Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea Molecular and Cellular Probes Portulaca oleracea Bacopa monnieri Diagnostic PCR Traditional Chinese medicine (TCM) Authentication |
| title | Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea |
| title_full | Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea |
| title_fullStr | Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea |
| title_full_unstemmed | Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea |
| title_short | Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea |
| title_sort | development of a specific and sensitive diagnostic pcr for rapid molecular authentication of the medicinal plant portulaca oleracea |
| topic | Portulaca oleracea Bacopa monnieri Diagnostic PCR Traditional Chinese medicine (TCM) Authentication |
| url | http://www.sciencedirect.com/science/article/pii/S0890850822001013 |
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