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Study on molecular identification methods and reagents “genetic marker”-based DNA fingerprinting-test strips of human placentophagy
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Yan-shuang WANG1, 3, Yi-tong WANG2, Run-hong MU1, **, Li-hua ZHANG4
Chinese Journal of Pharmaceutical Analysis | 2024, 44(7) : 1276 - 1284
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Chinese Journal of Pharmaceutical Analysis | 2024, 44(7): 1276-1284
Rapid Analys
Study on molecular identification methods and reagents “genetic marker”-based DNA fingerprinting-test strips of human placentophagy
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Yan-shuang WANG1, 3, Yi-tong WANG2, Run-hong MU1, **, Li-hua ZHANG4
Affiliations
  • 1.Basic Medicine School, Beihua University, Jilin 132013, China
  • 2.Second Hospital, Jilin University, Changchun 130000, China
  • 3.Chinese Medicine DNA Fingerprint Detection Technology Technology Innovation Center, Beihua University, Jilin 132013, China
  • 4.Jilin Leining Food and Drug Testing Technology Service Co., Ltd., Jilin 132013, China
Published: 2024-07-31 doi: 10.16155/j.0254-1793.2023-0435
Outline
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Objective:

To establish a rapid molecular identification method for DNA fingerprinting-immunocolloidal gold test strips of Chinese herbal medicine human placentophagy genetic markers and to develop an integrated quality control gene detection kit.

Methods:

Self-developed method and reagents could be for template DNA extraction from human placentophagy. Species-specific primers were designed according to the human mtDNA cytb gene, and the 5’ ends of the primers were labeled with FAM and Biotin, respectively. The optimal annealing temperature and cycle number of PCR were screened to establish a DNA fingerprinting molecular identification method. To develop PCR gene detection reagent premixes, to prepare DNA clones of control herbs using molecular cloning and gene sequencing techniques and to utilize immunocolloidal gold test strips for result detection. Finally, a rapid gene detection kit integrating DNA extraction reagent, PCR gene detection reagent premixes, control herb DNA clone solution, blank control solution and test strips was developed, and evaluated for specificity, reproducibility, sensitivity and stability.

Results:

The template DNA extracted by self-developed DNA extraction reagents were all in the concentration of 150-280 ng·μL-1, the purities were all in the range of 1.8-1.9, and there were no trail in the agarose gel electrophoresis. And the immunocolloidal gold test strips showed two red bands for human placentophagy control herb and authentic product, and one red band for easy-mix product and blank control at annealing temperature of 59 ℃ and 20 cycles for human placentophagy specific primers. The agarose gel electrophoresis was verified to be correct. The DNA sequencing results of the control herbs were 100% homologous with human mtDNA cytb gene. The self-developed integrated rapid gene detection kit was specific, reproducible, stable and sensitive at 0.1 ng·μL-1.

Conclusion:

The DNA fingerprinting-immunocolloidal gold test strip molecular identification method can specifically, accurately, rapidly and visually identify the authenticity of human placentophagy, and the integrated rapid gene detection kit provides a regulated and standardized testing method for quality control of human placentophagy. Therefore, it is more suitable for on-site field testing and universal promotion.

Placenta Hominis  /  genetic marker  /  mitochondrial cytochrome b  /  PCR technique  /  DNA fingerprinting  /  immunocolloidal gold test strip  /  integrated reagent kit
Yan-shuang WANG, Yi-tong WANG, Run-hong MU, Li-hua ZHANG. Study on molecular identification methods and reagents “genetic marker”-based DNA fingerprinting-test strips of human placentophagy[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (7) : 1276 -1284 . DOI: 10.16155/j.0254-1793.2023-0435
Year 2024 volume 44 Issue 7
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Article Info
doi: 10.16155/j.0254-1793.2023-0435
  • Online Date:2026-03-13
  • Published:2024-07-31
Article Data
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History
  • Revised:2024-05-30
Funding
Affiliations
    1.Basic Medicine School, Beihua University, Jilin 132013, China
    2.Second Hospital, Jilin University, Changchun 130000, China
    3.Chinese Medicine DNA Fingerprint Detection Technology Technology Innovation Center, Beihua University, Jilin 132013, China
    4.Jilin Leining Food and Drug Testing Technology Service 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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