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Establishment and evaluation of a rapid PCR-colloidal gold test strip method for the detection of Fritillaria ussuriensis
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Yu-he MA1, Cong-hui SHANG1, Qiu-he MA1, Tao LI1, Yue LIU1, Bei-zhen PAN1, Li-jun GAO1, *, Ming-cheng LI1, 2, Wei XIA1, Yong-mei QU3
Acta Pharmaceutica Sinica | 2024, 59(6) : 1773 - 1778
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Acta Pharmaceutica Sinica | 2024, 59(6): 1773-1778
Original Articles
Establishment and evaluation of a rapid PCR-colloidal gold test strip method for the detection of Fritillaria ussuriensis
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Yu-he MA1, Cong-hui SHANG1, Qiu-he MA1, Tao LI1, Yue LIU1, Bei-zhen PAN1, Li-jun GAO1, *, Ming-cheng LI1, 2, Wei XIA1, Yong-mei QU3
Affiliations
  • 1. School of Medical Technology, Beihua University, Jilin 132013, China
  • 2. Innovation Center for Detection on DNA Fingerprint of Traditional Chinese Medicine, Jilin 132013, China
  • 3. Jilin Guoan Pharmaceutical Co., Ltd., Jilin 132013, China
Published: 2024-06-12 doi: 10.16438/j.0513-4870.2023-1355
Outline
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This study design of specific identification primers for the ITS2 sequence of F. ussuriensis. The reaction system and conditions were optimized, and PCR-nucleic acid test strips were constructed to realize the visual detection of F. ussuriensis Bark. Through molecular cloning and sequencing technology, we constructed a positive control for F. ussuriensis DNA and formulated quality standards. The established method was evaluated for sensitivity, specificity and reproducibility, and the authenticity of the commercially available samples was identified. Results demonstrated that based on the ITS2 sequences, F. ussuriensis and its mixed forgeries could be distinguished. The PCR products of the authentic F. ussuriensis on test strips showed two bands in the T and C lines, while the pseudo products and negative control showed only one band in the C line, which was consistent with the results of agarose gel electrophoresis. The specificity was 100%, and the sensitivity of the PCR-nucleic acid test strip was up to 0.1 ng·μL-1, which was 10 times higher than that of the gel electrophoresis assay. 11 out of 16 commercially available samples of F. ussuriensis were qualified, and 5 were unqualified. Collectively, the PCR-nucleic acid test strip method established in this study is specific, rapid, accurate and visualized, which can provide a new technical idea for the detection of F. ussuriensis.

F. ussuriensis  /  molecular cloning  /  ITS2 sequence  /  nucleic acid test strip  /  visualization
Yu-he MA, Cong-hui SHANG, Qiu-he MA, Tao LI, Yue LIU, Bei-zhen PAN, Li-jun GAO, Ming-cheng LI, Wei XIA, Yong-mei QU. Establishment and evaluation of a rapid PCR-colloidal gold test strip method for the detection of Fritillaria ussuriensis[J]. Acta Pharmaceutica Sinica, 2024 , 59 (6) : 1773 -1778 . DOI: 10.16438/j.0513-4870.2023-1355
Year 2024 volume 59 Issue 6
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Article Info
doi: 10.16438/j.0513-4870.2023-1355
  • Receive Date:2023-12-02
  • Online Date:2025-11-26
  • Published:2024-06-12
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History
  • Received:2023-12-02
  • Revised:2024-02-28
Funding
Affiliations
    1. School of Medical Technology, Beihua University, Jilin 132013, China
    2. Innovation Center for Detection on DNA Fingerprint of Traditional Chinese Medicine, Jilin 132013, China
    3. Jilin Guoan Pharmaceutical Co., Ltd., Jilin 132013, China
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表12种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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